Peptides and peptide mimetics to treat pathologies associated with eye disease

ABSTRACT

This invention provides novel active agents (e.g. peptides, small organic molecules, amino acid pairs, etc.) peptides that ameliorate one or more symptoms of eye disease and/or other pathologies characterized by an inflammatory response, hi certain embodiment, the peptides resemble a G* amphipathic helix of apolipoprotein J. The agents are highly stable and readily administered via an oral route or via intraocular injection.

CROSS-REFERENCE TO RELATED PATENT APPLICATIONS

This application claims priority to U.S. application Ser. No.12/027,728, titled Peptides and Peptide Mimetics to Treat PathologiesAssociated with Eye Disease, filed on Feb. 7, 2008, which isincorporated herein by reference in their entirety for all purposes.

STATEMENT AS TO RIGHTS TO INVENTIONS MADE UNDER FEDERALLY SPONSOREDRESEARCH AND DEVELOPMENT

This invention was made with government support under Grant No: HL30568and Grant No: ey 06109 awarded by the National Heart Blood LungInstitute of the National Institutes of Health and the National EyeInstitute. The Government of the United States of America has certainrights in the invention.

FIELD OF THE INVENTION

This invention relates to the field of atherosclerosis and otherconditions characterized by inflammation and/or the formation of variousoxidized species. In particular, this invention pertains to theidentification of classes of active agents that are orally administrableand that ameliorate one or more symptoms of conditions characterized byan inflammatory response and/or the formation of various oxidizedspecies. This invention also relates to the field of maculardegeneration. In particular, this invention pertains to methods oftreating macular degeneration as well as methods of ameliorating asymptom of macular degeneration.

BACKGROUND OF THE INVENTION

The introduction of statins (e.g., Mevacor®, Lipitor®, etc.) has reducedmortality from heart attack and stroke by about one-third. However,heart attack and stroke remain the major cause of death and disability,particularly in the United States and in Western European countries.Heart attack and stroke are the result of a chronic inflammatorycondition, which is called atherosclerosis.

Several causative factors are implicated in the development ofcardiovascular disease including hereditary predisposition to thedisease, gender, lifestyle factors such as smoking and diet, age,hypertension, and hyperlipidemia, including hypercholesterolemia.Several of these factors, particularly hyperlipidemia andhypercholesteremia (high blood cholesterol concentrations) provide asignificant risk factor associated with atherosclerosis.

Cholesterol is present in the blood as free and esterified cholesterolwithin lipoprotein particles, commonly known as chylomicrons, very lowdensity lipoproteins (VLDLs), low density lipoproteins (LDLs), and highdensity lipoproteins (HDLs). Concentration of total cholesterol in theblood is influenced by (1) absorption of cholesterol from the digestivetract, (2) synthesis of cholesterol from dietary constituents such ascarbohydrates, proteins, fats and ethanol, and (3) removal ofcholesterol from blood by tissues, especially the liver, and subsequentconversion of the cholesterol to bile acids, steroid hormones, andbiliary cholesterol.

Maintenance of blood cholesterol concentrations is influenced by bothgenetic and environmental factors. Genetic factors include concentrationof rate-limiting enzymes in cholesterol biosynthesis, concentration ofreceptors for low density lipoproteins in the liver, concentration ofrate-limiting enzymes for conversion of cholesterols bile acids, ratesof synthesis and secretion of lipoproteins and gender of person.Environmental factors influencing the hemostasis of blood cholesterolconcentration in humans include dietary composition, incidence ofsmoking, physical activity, and use of a variety of pharmaceuticalagents. Dietary variables include the amount and type of fat (saturatedand polyunsaturated fatty acids), the amount of cholesterol, amount andtype of fiber, and perhaps the amounts of vitamins such as vitamin C andD and minerals such as calcium.

Low density lipoprotein (LDL) oxidation has been strongly implicated inthe pathogenesis of atherosclerosis. High density lipoprotein (HDL) hasbeen found to be capable of protecting against LDL oxidation, but insome instances has been found to accelerate LDL oxidation. Importantinitiating factors in atherosclerosis include the production ofLDL-derived oxidized phospholipids.

Normal HDL has the capacity to prevent the formation of these oxidizedphospholipids and also to inactivate these oxidized phospholipids oncethey have formed. However, under some circumstances HDL can be convertedfrom an anti-inflammatory molecule to a pro-inflammatory molecule thatactually promotes the formation of these oxidized phospholipids.

It has been suggested that HDL and LDL function as part of the innateimmune system (Navab et al. (2001) Arterioscler. Thromb. Vasc. Biol.,21: 481-488). The generation of anti-inflammatory HDL has been achievedusing class A amphipathic helical peptides that mimic the major proteinof HDL, apolipoprotein A-I (apo A-I) (see, e.g., WO 02/15923).

Age-related macular degeneration (AMD) is the most frequent cause oflegal blindness in the elderly in industrial countries (Van Leeuwen etal. (2003) European Journal of Epidemiology 18: 845-854). It is aheterogeneous disease, which is characterized by progressive loss ofcentral, high acuity vision. For the patient it compromises dramaticallyquality of life, since they lose their ability to read, to recognizefaces and day-to-day tasks become major obstacles. According to the WHOa total of 30-50 million individuals are affected and about 14 millionpeople are blind or severely visually impaired due to AMD (Gehrs et al.,(2006) Annals of Medicine 38:450-471).

The most prominent clinical and histopathological lesions of AMD involvethe choriocapillaris, Bruch's membrane, and the retinal pigmentepithelium (RPE) (Ambati et al. (2003) Survey of Opthalmology48:257-293). The choriocapillaris is a highly specialized capillaryplexus with the highest blood flow rate in the body which interacts withthe highly metabolic active RPE. The RPE forms the outer blood-retinabarrier and supplies the photoreceptors, the sensory cells in the eye,with nutriments as well as phagocytes daily shed outer photoreceptorsegments which are degraded and partially recycled. Under normalconditions unrecycled end products are rendered into thechoriocapillaris. Bruch's membrane is a five layer connective tissuebetween the RPE and choriocapillaris resembling an arterial intima inits function (Curcio et al. (2001) Invest Opthalmol Vis Sci 42:265-274).With age Bruch's membrane undergoes distinctive degenerative changes.One major characteristic feature next to thickening is the accumulationof neutral lipids, which build up a diffusion barrier between the RPEand choriocapillaris compromising RPE and photoreceptor function (Curcioet al. (2001) Invest Opthalmol Vis Sci 42:265-274; Pauleikhoff et al.(1990) Opthalmology 97:171-178; Moore et al. (1995) Invest Opthalmol VisSci 36:1290-1297).

In early stages of AMD an additional deposition of debris is observedbetween the basal membrane of the RPE (1^(st) layer of Bruch's membrane)and the inner collagenous layer (2^(nd) layer of Bruch's membrane). Thisdebris is called basal linear deposits and drusen, both rich in lipidsand hallmarks of AMD, impairing even more the diffusion along Bruch'smembrane (Gehrs et al, (2006) Annals of Medicine 38:450-471; Curcio etal. (1999) Arch Opthalmol 117:329-339; Curcio et al. (2005) ExperimentalEye Research 81: 731-741; Haimovici et al. (2001) Invest Opthalmol VisSci 42:1592-1599). Furthermore, cytotoxic and lipid rich, metabolic endproducts, called lipofuscin, accumulate in the RPE cells (Beatty et al.(2000) Surv Opthalmol 45:115-134). All these conditions together causeoxidative stress and inflammation resulting in RPE atrophy andsuccessively photoreceptor degeneration (Kopitz et al. (2004) Biochimie86: 825-831). This atrophy of RPE and photoreceptors is called the dryform of AMD and progresses slowly and irreversibly. Currently atreatment or prevention of this form of AMD, which affect about 85-90%of all AMD patients, does not exist (Van Leeuwen et al. (2003) EuropeanJournal of Epidemiology 18: 845-854).

The second form of AMD is called wet AMD and can arise from the dryform. It affects about 10-15% of all AMD patients and is marked by thegrowth of a pathological vessel from the choriocapillaris into thesubretinal space, called choroidal neovascularization (CNV) (Gehrs etal. (2006) Annals of Medicine 38:450-471 and Ambati et al. (2003) Surveyof Opthalmology 48:257-293). It causes a rapid, irreversible vision lossdue to leakage, bleeding, and scaring (Ambati et al. (2003) Survey ofOpthalmology 48:257-293). In the last 5 years antiangiogentic therapieswere developed targeting vascular endothelial growth factor, which couldshow success in slowing down the progression of vision loss (Michels etal. (2006) Expert Opin Investig Drugs 15:779-793).

In general, current therapies use antibodies or antibody fragmentsagainst VEGF, which are injected into the vitreous body of the eye(Michels et al. (2006)). A prevention therapy of wet AMD does not exist(Gehrs et al. (2006)), which would be especially desirable when thevision in one eye is already largely compromised and the second eyeshows definite risk factors for a progression like e.g. large softdrusen (Ambati et al. (2003)).

Lipids are hydrophobic and cannot simply dissolve in an aqueous mediumsuch as blood. In order to be transported in blood lipids have to beassembled in particles called lipoproteins. Specialized proteins calledapolipoproteins help to form and stabilize these particles. There areseveral classes of apolipoproteins (A-JM, a). Basically their functionalstructures are comparable which are amphipathic helices.

Apolipoprotein mimetic peptides are synthetic helical lipid acceptingpeptides mimicking the function of an apolipoprotein (Mendez et al.(1994) J. Clin. Invest 94:1698-1705). One of the best known is theApoA-I mimetic peptide 4F, has been shown to treat atherosclerosis(Anantharamaiah et al. (2006) Current Opinion in Lipidology 17:233-237).It is available as L-4F and as its stereoisomer D-4F. It consists of 18amino acids, is well water-soluble, a high potent lipid acceptor, andacts as a highly anti-inflammatory (Navab et al. (2006) Nat. Clin.Pract. Cardiovasc. Med. 3:540-547). D-4F is based on D-amino acids andis compared to L-4F more resistant to degradation and can be takenorally (Anantharamaiah et al. (2006)). A phase I clinical trial withD-4F already started. So far no side effects of D-4F are described.

SUMMARY OF THE INVENTION

This invention provides novel compositions and methods to ameliorate oneor more symptoms of a vascular condition and/or a conditioncharacterized by an inflammatory response and/or a conditioncharacterized by the formation of oxidized reactive species in a mammal.

Thus, in certain embodiments, this invention provides a peptide thatameliorates a symptom of atherosclerosis, where the peptide comprisesthe amino acid sequence or the retro amino acid sequence of a peptidelisted in Table 6. In another embodiment this invention provides apeptide that ameliorates a symptom of atherosclerosis, where thepeptide: consists of 18 amino acids, the 18 amino acids consisting of 3alanines (A), 2 aspartates (D), 2 glutamates (E), 4 phenylalanines (F),4 lysines (K), 1 valine (V), 1 tryptophan (W), and 1 tyrosine (Y); wherethe peptide forms a class A amphipathic helix; comprises at least one“D” amino acid residue; and protects a phospholipid against oxidation byan oxidizing agent. In certain embodiments these peptides include butare not limited to a peptide having the amino acid sequence or the retroamino acid sequence of a peptide listed in Table 4. In still anotherembodiment, this invention provides a peptide that ameliorates a symptomof atherosclerosis, where the peptide: ranges in length from about 18 to37 amino acids and comprises at least 3 alanines (A), 2 aspartates (D),2 glutamates (E), 4 phenylalanines (F), 4 lysines (K), 1 valine (V), 1tryptophan (W), 1 tyrosine (Y); where the peptide forms a class Aamphipathic helix; comprises at least one “D” amino acid residue; andprotects a phospholipid against oxidation by an oxidizing agent. Incertain embodiments these peptides comprise an amino acid sequenceselected from the group consisting ofD-W-F-K-A-F-Y-D-K-V-A-E-K-F-K-E-A-F (SEQ ID NO: 1191),-D-W-L-K-A-F-Y-D-K-V-A-E-K-L-K-E-A-F-P-D-W-L-K-A-F-Y-D-K-V-A-E-K-L-K-E-A-F(SEQ ID NO: 1192),-D-W-L-K-A-F-Y-D-K-V-A-E-K-L-K-E-F-F-P-D-W-L-K-A-F-Y-D-K-V-A-E-K-L-K-E-F-F(SEQ ID NO: 1193),-D-W-F-K-A-F-Y-D-K-V-A-E-K-L-K-E-A-F-P-D-W-F-K-A-F-Y-D-K-V-A-E-K-L-K-E-A-F(SEQ ID NO: 1194),D-K-L-K-A-F-Y-D-K-V-F-E-W-A-K-E-A-F-P-D-K-L-K-A-F-Y-D-K-V-F-E-W-L-K-E-A-F(SEQ ID NO: 1195),D-K-W-K-A-V-Y-D-K-F-A-E-A-F-K-E-F-L-P-D-K-W-K-A-V-Y-D-K-F-A-E-A-F-K-E-F-L(SEQ ID NO: 1196),D-W-F-K-A-F-Y-D-K-V-A-E-K-F-K-E-A-F-P-D-W-F-K-A-F-Y-D-K-V-A-E-K-F-K-E-A-F-(SEQID NO: 1197), or the reverse of any of these sequences. In still yetanother embodiment this invention provides a peptide that forms a classA amphipathic helix or a class Y amphipathic helix and is described bythe formula: D¹-X¹-X¹-K¹-Y¹-X³-X⁴-D²-K²-X-⁵-Y-D³-K³-X⁶-K⁴-D⁴-Y²-X⁷ whereX¹, X², X³, X⁴, X⁵, and X⁶ are independently selected from the groupconsisting of Leu, norLeu, Val, Ile, Trp, Phe, Tyr, β-Nal, and α-Nal,and all X residues are on the non-polar face of the peptide, except forone that can be on the polar face between two K residues; K′, K², K³,and K⁴ are independently Lys or Arg, and no more than two K's areadjacent to each other in a helical wheel diagram of the peptide; Y¹ andY² are independently selected from the group consisting of Ala, His,Ser, Gln, Asn, and Thr, when present on the non-polar face of themolecule; when one of Y¹ or Y² are present on the polar face of themolecule, the Y¹ or Y² on the polar face of the molecule is selectedfrom the group consisting of Ala, His, Ser, Gln, Asn, and Thr; D¹, D²,D³, and D⁴ are independently Asp or Glu, and no more than 3 Ds arecontiguous in a helical wheel diagram of the peptide, and the remainingD is separated from the other D's by a Y. In certain embodiments thesepeptides comprise the amino acid sequence or the retro amino acidsequence of a peptide listed in Table 5.

In certain embodiments any one or more of these peptides furthercomprise a protecting group coupled to the amino or carboxyl terminus.In certain embodiments the peptides comprise a first protecting groupcoupled to the amino terminus and a second protecting group coupled tothe carboxyl terminus. In certain embodiments the protecting groups canbe independently selected from the group consisting of acetyl, amide,and 3 to 20 carbon alkyl groups, Fmoc, Tboc, 9-fluoreneacetyl group,1-fluorenecarboxylic group, 9-florenecarboxylic group,9-fluorenone-1-carboxylic group, benzyloxycarbonyl, Xanthyl (Xan),Trityl (Trt), 4-methyltrityl (Mtt), 4-methoxytrityl (Mmt),4-methoxy-2,3,6-trimethyl-benzenesulphonyl (Mtr), Mesitylene-2-sulphonyl(Mts), 4,4-dimethoxybenzhydryl (Mbh), Tosyl (Tos), 2,2,5,7,8-pentamethylchroman-6-sulphonyl (Pmc), 4-methylbenzyl (MeBzl), 4-methoxybenzyl(MeOBzl), Benzyloxy (BzlO), Benzyl (Bzl), Benzoyl (Bz),3-nitro-2-pyridinesulphenyl (Npys),1-(4,4-dimentyl-2,6-diaxocyclohexylidene)ethyl (Dde), 2,6-dichlorobenzyl(2,6-DiCl-Bzl), 2-chlorobenzyloxycarbonyl (2—Cl-Z),2-bromobenzyloxycarbonyl (2—Br-Z), Benzyloxymethyl (Bom),t-butoxycarbonyl (Boc), cyclohexyloxy (cHxO), t-butoxymethyl (Bum),t-butoxy (tBuO), t-Butyl (tBu), Acetyl (Ac), and Trifluoroacetyl (TFA).

In certain embodiments the peptide comprises a protecting group coupledto the amino terminal and the amino terminal protecting group is aprotecting group selected from the group consisting of acetyl,propeonyl, and a 3 to 20 carbon alkyl. In certain embodiments thepeptide comprises a protecting group coupled to the carboxyl terminaland the carboxyl terminal protecting group is an amide. In certainembodiments the peptide comprises: a first protecting group coupled tothe amino terminus where the protecting group is a protecting groupselected from the group consisting of acetyl, propeonyl, and a 3 to 20carbon alkyl; and a second protecting group coupled to the carboxylterminal and the carboxyl terminal protecting group is an amide.

In various embodiments one or more amino acids comprising the peptideare “D” amino acids. In various embodiments all amino acids comprisingthe peptide “D” amino acids. The peptide(s) can, optionally, bemixed/combined with a pharmacologically acceptable excipient. In certainembodiments the excipient is an excipient suitable for oraladministration to a mammal.

In certain embodiments this invention provides methods of treating avascular condition and/or a condition characterized by an inflammatoryresponse and/or a condition characterized by the formation of oxidizedreactive species in a mammal. The methods typically involveadministering to a mammal in need thereof one or more of the activeagents described in Tables 2-18, and/or a small organic molecule asdescribed herein in an amount sufficient to ameliorate one or moresymptoms of the condition. In certain embodiments the active agent is apolypeptide comprising the amino acid sequence of 4F (SEQ ID NO:5). Incertain embodiments the administration is by a route selected from thegroup consisting of oral administration, nasal administration, rectaladministration, intraperitoneal injection, and intravascular injection,intraocular injection, intravitreal injection, subconjuctival injection,peri-/retrobulbar injection, subcutaneous injection, eye drops, eye gel,eye ointment, spray, emulsion, suspension, transcutaneousadministration, and intramuscular injection, via any drug carriers assponges, contact lenses, polymers, microspheres, implants, pellets, andgenetically engineered cells. In certain embodiments the active agent isadministered in conjunction with a drug selected from the groupconsisting of CETP inhibitors, FTY720, Certican, DPP4 inhibitors,Calcium channel blockers, ApoA1 derivative or mimetic or agonist, PPARagonists, Steroids, Gleevec, Cholesterol Absorption blockers (Zetia),Vytorin, Any Renin Angiotensin pathway blockers, Angiotensin II receptorantagonist (Diovan etc), ACE inhibitors, Renin inhibitors, MR antagonistand Aldosterone synthase inhibitor, Beta-blockers, Alpha-adrenergicantagonists, LXR agonist, FXR agonist, Scavenger Receptor B1 agonist,ABCA1 agonist, Adiponectic receptor agonist or adiponectin inducers,Stearoyl-CoA Desaturase I (SCD1) inhibitor, Cholesterol synthesisinhibitors (non-statins), Diacylglycerol Acyltransferase I (DGAT1)inhibitor, Acetyl CoA Carboxylase 2 inhibitor, PAI-1 inhibitor, LP-PLA2inhibitor, GLP-1, Glucokinase activator, CB-1 agonist, AGEinhibitor/breaker, PKC inhibitors, Anti-thrombotic/coagulants: Aspirin,ADP receptor blockers e.g. Clopidigrel, Factor Xa inhibitor, GPIIb/IIIainhibitor, Factor VIIa inhibitor, Warfarin, Low molecular weightheparin, Tissue factor inhibitor, Anti-inflammatory drugs: Probucol andderivative e.g. AGI-1067 etc, CCR2 antagonist, CX3CR1 antagonist, IL-1antagonist, Nitrates and NO donors, and Phosphodiesterase inhibitors.

In various embodiments this invention provides for the use of an activeagent described in Tables 2-18, and/or a small organic molecule asdescribed herein in a treatment of a condition selected from the groupconsisting of atherosclerotic plaque formation, atherosclerotic lesionformation, myocardial infarction, stroke, congestive heart failure,arteriole function, arteriolar disease, arteriolar disease associatedwith aging, arteriolar disease associated with Alzheimer's disease,arteriolar disease associated with chronic kidney disease, arteriolardisease associated with hypertension, arteriolar disease associated withmulti-infarct dementia, arteriolar disease associated with subarachnoidhemorrhage, peripheral vascular disease, chronic obstructive pulmonarydisease (COPD), emphysema, asthma, idiopathic pulmonary fibrosis,pulmonary fibrosis, adult respiratory distress syndrome, osteoporosis,Paget's disease, coronary calcification, rheumatoid arthritis,polyarteritis nodosa, polymyalgia rheumatica, lupus erythematosus,multiple sclerosis, Wegener's granulomatosis, central nervous systemvasculitis (CNSV), Sjogren's syndrome, scleroderma, polymyositis, AIDSinflammatory response, bacterial infection, fungal infection, viralinfection, parasitic infection, influenza, avian flu, viral pneumonia,endotoxic shock syndrome, sepsis, sepsis syndrome, trauma/wound, organtransplant, transplant atherosclerosis, transplant rejection, cornealulcer, chronic/non-healing wound, ulcerative colitis, reperfusion injury(prevent and/or treat), ischemic reperfusion injury (prevent and/ortreat), spinal cord injuries (mitigating effects), cancers,myeloma/multiple myeloma, ovarian cancer, breast cancer, colon cancer,bone cancer, osteoarthritis, inflammatory bowel disease, allergicrhinitis, cachexia, diabetes, Alzheimer's disease, implanted prosthesis,biofilm formation, Crohns' disease, dermatitis, acute and chronic,eczema, psoriasis, contact dermatitis, scleroderma, Type I Diabetes,Type II Diabetes, juvenile onset diabetes, prevention of the onset ofdiabetes, diabetic nephropathy, diabetic neuropathy, diabeticretinopathy, erectile dysfunction, macular degeneration, multiplesclerosis, nephropathy, neuropathy, Parkinson's Disease, peripheralvascular disease, and meningitis.

This invention additionally provides for the use of active agentdescribed in Tables 2-18, and/or a small organic molecule as describedherein for the manufacture of a medicament for the treatment of acondition selected from the group consisting of atherosclerotic plaqueformation, atherosclerotic lesion formation, myocardial infarction,stroke, congestive heart failure, arteriole function, arteriolardisease, arteriolar disease associated with aging, arteriolar diseaseassociated with Alzheimer's disease, arteriolar disease associated withchronic kidney disease, arteriolar disease associated with hypertension,arteriolar disease associated with multi-infarct dementia, arteriolardisease associated with subarachnoid hemorrhage, peripheral vasculardisease, chronic obstructive pulmonary disease (COPD), emphysema,asthma, idiopathic pulmonary fibrosis, pulmonary fibrosis, adultrespiratory distress syndrome, osteoporosis, Paget's disease, coronarycalcification, rheumatoid arthritis, polyarteritis nodosa, polymyalgiarheumatica, lupus erythematosus, multiple sclerosis, Wegener'sgranulomatosis, central nervous system vasculitis (CNSV), Sjogren'ssyndrome, scleroderma, polymyositis, AIDS inflammatory response,bacterial infection, fungal infection, viral infection, parasiticinfection, influenza, avian flu, viral pneumonia, endotoxic shocksyndrome, sepsis, sepsis syndrome, trauma/wound, organ transplant,transplant atherosclerosis, transplant rejection, corneal ulcer,chronic/non-healing wound, ulcerative colitis, reperfusion injury(prevent and/or treat), ischemic reperfusion injury (prevent and/ortreat), spinal cord injuries (mitigating effects), cancers,myeloma/multiple myeloma, ovarian cancer, breast cancer, colon cancer,bone cancer osteoarthritis, inflammatory bowel disease, allergicrhinitis, cachexia, diabetes, Alzheimer's disease, implanted prosthesis,biofilm formation, Crohns' disease, dermatitis, acute and chronic,eczema, psoriasis, contact dermatitis, scleroderma, Type I Diabetes,Type II Diabetes, juvenile onset diabetes, prevention of the onset ofdiabetes, diabetic nephropathy, diabetic neuropathy, diabeticretinopathy, erectile dysfunction, macular degeneration, multiplesclerosis, nephropathy, neuropathy, Parkinson's Disease, peripheralvascular disease, and meningitis.

In certain embodiments this invention provides a stent for deliveringdrugs to a vessel in a body. The stent typically comprises a stentframework including a plurality of reservoirs formed therein, and apeptide comprising the amino acid sequence or the retro amino acidsequence of a peptide listed in Tables 2-18 (e.g., Table 4, Table 5, orTable 6) and/or the inverse thereof. In certain embodiments the stentcomprises a peptide comprising the amino acid sequence of 4F (SEQ IDNO:5) or the inverse thereof. In certain embodiments the active agent iscontained within a polymer. In certain embodiments the stent frameworkcomprises one of a metallic base or a polymeric base. In certainembodiments the stent framework base comprises a material selected fromthe group consisting of stainless steel, nitinol, tantalum, MP35N alloy,platinum, titanium, a suitable biocompatible alloy, a suitablebiocompatible polymer, and a combination thereof. The reservoir(s)comprising said stent can, in some embodiments, comprise micropores(e.g. having a diameter of about 20 microns or less). In certainembodiments the micropores have a diameter in the range of about 20microns to about 50 microns. In various embodiments the micropores havea depth in the range of about 10 to about 50 microns. The micropores, incertain embodiments, extend through the stent framework having anopening on an interior surface of the stent and an opening on anexterior surface of the stent. In various embodiments the stent canfurther comprise a cap layer disposed on the interior surface of thestent framework, the cap layer covering at least a portion of thethrough-holes and providing a barrier characteristic to control anelution rate of a drug in the drug polymer from the interior surface ofthe stent framework. In various embodiments the reservoirs comprisechannels along an exterior surface of the stent framework. In variousembodiments the polymer comprises a first layer of a first drug polymerhaving a first pharmaceutical characteristic and the polymer layercomprises a second drug polymer having a second pharmaceuticalcharacteristic. In certain embodiments the stent further comprises abarrier layer positioned between the polymer comprising the activeagent. In various embodiments a catheter can be coupled to the stentframework. In certain embodiments the catheter can include a balloonused to expand the stent. In certain embodiments the catheter includes asheath that retracts to allow expansion of the stent.

Also provided is a method of manufacturing a drug-polymer stent. Themethod typically involves providing a stent framework; cutting aplurality of reservoirs in the stent framework; applying a compositioncomprising one or more peptides comprising the amino acid sequence orthe retro amino acid sequence of a peptide listed in any of Tables 2-18to at least one reservoir; and drying the composition. The method canfurther involve applying a polymer layer to the dried composition; anddrying the polymer layer.

This invention also provides a method of treating a vascular condition.The method involves positioning a stent as described above, within avessel of a body; expanding the stent; and eluting at least one activeagent (e.g., an active agent from any of Tables 2-18) from at least asurface of the stent.

In certain embodiments, this invention expressly excludes one or more ofthe peptides described in U.S. Pat. Nos. 6,037,323; 4,643,988;6,933,279; 6,930,085; 6,664,230; 3,767,040; 6,037,323; U.S. PatentPublications 2005/0164950; 2004/0266671; 2004/0254120; 2004/0057871;2003/0229015; 2003/0191057; 2003/0171277; 2003/0045460; 2003/0040505;PCT Publications WO 2002/15923; WO 1999/16408; WO 1997/36927; and/or inGarber et al. (1992) Arteriosclerosis and Thrombosis, 12: 886-894, whichare incorporated herein by reference.

Also disclosed herein are methods for treating a subject with eyedisease, the method comprising administering to the subject in needthereof an effective amount of one or more of the active agentsdescribed in Tables 2-18, and/or a small organic molecule as describedherein in an amount sufficient to ameliorate one or more symptoms ofsaid condition. The active agent can be a polypeptide comprising theamino acid sequence of 4F (SEQ ID NO:5). Administration can be by aroute selected from the group consisting of oral administration, nasaladministration, rectal administration, intraperitoneal injection, andintravascular injection, intraocular injection, intravitreal injection,subconjuctival injection, peri-/retrobulbar injection, subcutaneousinjection, eye drops, eye gel, eye ointment, spray, emulsion,suspension, transcutaneous administration, and intramuscular injection,via any drug carriers as sponges, contact lenses, polymers,microspheres, implants, pellets, and genetically engineered cells.

Also disclosed herein are methods for treating a subject with eyedisease, the method comprising administering to the subject in needthereof an effective amount of one or more of the active agentsdescribed in Tables 2-18, and/or a small organic molecule as describedherein in an amount sufficient to ameliorate one or more symptoms ofsaid condition, wherein said active agent is administered in conjunctionwith an antiangiogenic agent.

Also disclosed herein are methods of ameliorating a symptom of eyedisease, the method comprising administering to the subject to thesubject in need thereof an effective amount of one or more of the activeagents described in Tables 2-18, and/or a small organic molecule asdescribed herein in an amount sufficient to ameliorate one or moresymptoms of said condition. Symptoms of eye disease can include, but arenot limited to accumulation of extracellular lipids in Bruch'smembranes, accumulation of lipid rich debris, vision loss, formation ofchoriocapillaris, thickening of the Bruch's membrane, accumulation ofneutral lipids in the Bruch's membrane, formation of a diffusion barrierbetween the retinal pigment epithelium and choriocapillaris, depositionof debris (basal linear deposits and drusen) between the basal membraneof the RPE, and the inner collagenous layer, accumulation of lipofuscinin the RPE cells, RPE atrophy, photoreceptor degeneration, choroidalneovascularization, as well as leakage, bleeding, scarring of the eye

Also disclosed herein are methods of ameliorating a symptom of eyedisease, the method comprising administering to the subject an effectiveamount of a peptide wherein said peptide: ranges in length from about 18to 37 amino acids and comprises at least 3 alanines (A), 2 aspartates(D), 2 glutamates (E), 4 phenylalanines (F), 4 lysines (K), 1 valine(V), 1 tryptophan (W), 1 tyrosine (Y); wherein said peptide forms aclass A amphipathic helix; comprises at least one “D” amino acidresidue; and protects a phospholipid against oxidation by an oxidizingagent.

Also disclosed herein are methods of ameliorating a symptom of eyedisease, the method comprising administering to the subject an effectiveamount of a peptide wherein said peptide: ranges in length from about 18to 37 amino acids and comprises at least 3 alanines (A), 2 aspartates(D), 2 glutamates (E), 4 phenylalanines (F), 4 lysines (K), 1 valine(V), 1 tryptophan (W), 1 tyrosine (Y); wherein said peptide forms aclass A amphipathic helix; comprises at least one “D” amino acidresidue; and protects a phospholipid against oxidation by an oxidizingagent, wherein said peptide further comprises a protecting group coupledto the amino or carboxyl terminus.

Also disclosed herein are methods of ameliorating a symptom of eyedisease, the method comprising administering to the subject an effectiveamount of a peptide wherein said peptide: ranges in length from about 18to 37 amino acids and comprises at least 3 alanines (A), 2 aspartates(D), 2 glutamates (E), 4 phenylalanines (F), 4 lysines (K), 1 valine(V), 1 tryptophan (W), 1 tyrosine (Y); wherein said peptide forms aclass A amphipathic helix; comprises at least one “D” amino acidresidue; and protects a phospholipid against oxidation by an oxidizingagent, wherein said peptide comprises an amino acid sequence selectedfrom the group consisting of D-W-F-K-A-F-Y-D-K-V-A-E-K-F-K-E-A-F (SEQ IDNO: 1191),-D-W-L-K-A-F-Y-D-K-V-A-E-K-L-K-E-A-F-P-D-W-L-K-A-F-Y-D-K-V-A-E-K-L-K-E-A-F(SEQ ID NO: 1192),-D-W-L-K-A-F-Y-D-K-V-A-E-K-L-K-E-F-F-P-D-W-L-K-A-F-Y-D-K-V-A-E-K-L-K-E-F-F(SEQ ID NO: 1193),-D-W-F-K-A-F-Y-D-K-V-A-E-K-L-K-E-A-F-P-D-W-F-K-A-F-Y-D-K-V-A-E-K-L-K-E-A-F(SEQ ID NO: 1194),D-K-L-K-A-F-Y-D-K-V-F-E-W-A-K-E-A-F-P-D-K-L-K-A-F-Y-D-K-V-F-E-W-L-K-E-A-F(SEQ ID NO: 1195),D-K-W-K-A-V-Y-D-K-F-A-E-A-F-K-E-F-L-P-D-K-W-K-A-V-Y-D-K-F-A-E-A-F-K-E-F-L(SEQ ID NO: 1196),D-W-F-K-A-F-Y-D-K-V-A-E-K-F-K-E-A-F-P-D-W-F-K-A-F-Y-D-K-V-A-E-K-F-K-E-A-F-(SEQID NO: 1197), or the reverse of any of these sequences.

Also discloses is the use of an active agent described in Tables 2-18,and/or a small organic molecule as described herein in a treatment ofmacular degeneration.

Also disclose are methods of treating a subject with eye disease, themethod comprising administering to the subject in need thereof aneffective amount of one or more of the active agents described in Tables2-18, and/or a small organic molecule as described herein in an amountsufficient to ameliorate one or more symptoms of said condition incombination with an anti-angiogenic therapy.

Also disclosed are methods of ameliorating a symptom of eye disease, themethod comprising administering to the subject to the subject in needthereof an effective amount of one or more of the active agentsdescribed in Tables 2-18, and/or a small organic molecule as describedherein in an amount sufficient to ameliorate one or more symptoms ofsaid condition in combination with an anti-angiogenic therapy.

DEFINITIONS

The term “treat” when used with reference to treating, e.g. a pathologyor disease refers to the mitigation and/or elimination of one or moresymptoms of that pathology or disease, and/or a reduction in the rate ofonset or severity of one or more symptoms of that pathology or disease,and/or the prevention of that pathology or disease.

The terms “isolated”, “purified”, or “biologically pure” when referringto an isolated polypeptide refer to material that is substantially oressentially free from components that normally accompany it as found inits native state. With respect to nucleic acids and/or polypeptides theterm can refer to nucleic acids or polypeptides that are no longerflanked by the sequences typically flanking them in nature. Chemicallysynthesized polypeptides are “isolated” because they are not found in anative state (e.g. in blood, serum, etc.). In certain embodiments, theterm “isolated” indicates that the polypeptide is not found in nature.

The terms “polypeptide”, “peptide” and “protein” are usedinterchangeably herein to refer to a polymer of amino acid residues. Theterms apply to amino acid polymers in which one or more amino acidresidues is an artificial chemical analogue of a corresponding naturallyoccurring amino acid, as well as to naturally occurring amino acidpolymers.

The term “an amphipathic helical peptide” refers to a peptide comprisingat least one amphipathic helix (amphipathic helical domain). Certainamphipathic helical peptides of this invention can comprise two or more(e.g., 3, 4, 5, etc.) amphipathic helices.

The term “class A amphipathic helix” refers to a protein structure thatforms an α-helix producing a segregation of a polar and nonpolar faceswith the positively charged residues residing at the polar-nonpolarinterface and the negatively charged residues residing at the center ofthe polar face (see, e.g., Segrest et al. (1990) Proteins: Structure,Function, and Genetics 8: 103-117).

“Apolipoprotein J” (apo J) is known by a variety of names includingclusterin, TRPM2, GP80, and SP 40 (see, e.g., Fritz (1995) Pp 112 In:Clusterin: Role in Vertebrate Development, Function, and Adaptation(Harmony JAK Ed.), R. G. Landes, Georgetown, Tex.,). It was firstdescribed as a heterodimeric glycoprotein and a component of thesecreted proteins of cultured rat Sertoli cells (see, e.g., Kissinger etal. (1982) Biol. Reprod.; 27: 233240). The translated product is asingle-chain precursor protein that undergoes intracellular cleavageinto a disulfide-linked 34 kDa α-subunit and a 47 kDa β-subunit (see,e.g., Collard and Griswold (1987) Biochem., 26: 3297-3303). It has beenassociated with cellular injury, lipid transport, apoptosis and it maybe involved in clearance of cellular debris caused by cell injury ordeath. Clusterin has been shown to bind to a variety of molecules withhigh affinity including lipids, peptides, and proteins and thehydrophobic probe 1-anilino-8-naphthalenesulfonate (Bailey et al. (2001)Biochem., 40: 11828-11840).

The class G amphipathic helix is found in globular proteins, and thus,the name class G. The feature of this class of amphipathic helix is thatit possesses a random distribution of positively charged and negativelycharged residues on the polar face with a narrow nonpolar face. Becauseof the narrow nonpolar face this class does not readily associate withphospholipid (see, e.g., Segrest et al. (1990) Proteins: Structure,Function, and Genetics. 8: 103-117; Erratum (1991) Proteins: Structure,Function and Genetics, 9: 79). Several exchangeable apolipoproteinspossess similar but not identical characteristics to the G amphipathichelix. Similar to the class G amphipathic helix, this other classpossesses a random distribution of positively and negatively chargedresidues on the polar face. However, in contrast to the class Gamphipathic helix which has a narrow nonpolar face, this class has awide nonpolar face that allows this class to readily bind phospholipidand the class is termed G* to differentiate it from the G class ofamphipathic helix (see, e.g., Segrest et al. (1992) J. Lipid Res., 33:141-166; Anantharamaiah et al. (1993) Pp. 109-142 In: The AmphipathicHelix, Epand, R. M. Ed CRC Press, Boca Raton, Fla.). Computer programsto identify and classify amphipathic helical domains have been describedby Jones et al. (1992) J. Lipid Res. 33: 287-296) and include, but arenot limited to the helical wheel program (WHEEL or WHEEL/SNORKEL),helical net program (HELNET, HELNET/SNORKEL, HELNET/Angle), program foraddition of helical wheels (COMBO or COMBO/SNORKEL), program foraddition of helical nets (COMNET, COMNET/SNORKEL, COMBO/SELECT,COMBO/NET), consensus wheel program (CONSENSUS, CONSENSUS/SNORKEL), andthe like.

The term “ameliorating” when used with respect to “ameliorating one ormore symptoms of atherosclerosis” refers to a reduction, prevention, orelimination of one or more symptoms characteristic of atherosclerosisand/or associated pathologies. Such a reduction includes, but is notlimited to a reduction or elimination of oxidized phospholipids, areduction in atherosclerotic plaque formation and rupture, a reductionin clinical events such as heart attack, angina, or stroke, a decreasein hypertension, a decrease in inflammatory protein biosynthesis,reduction in plasma cholesterol, and the like.

The term “eye disease” as used herein includes diseases associated orcompromised with/by a reduced hydraulic conductivity and metabolicexchange via Bruch's membrane, diseases characterized by an accumulationof extra-/intracellular lipids in the eye, diseases that uselipid-derived mediators of inflammation or benefit from oxidized lipidremoval as well as diseases that benefit from Bruch's membraneremodeling. “eye disease” as used herein includes, but is not limitedto, macular degeneration, age related maculopathy (ARM), age relatedmacular degeneration (AMD) including both the dry and wet forms of agerelated macular degeneration, glaucoma, ocular hypertension, macularedema, retinal pigment epithelium detachments, coats disease, uveitis,sicca syndrome, hereditary diseases associated with increasedextra-/intracellular lipid storage/accumulation, and juvenile maculardegeneration as well as all risk factors for each mentioned disease

The term “ameliorating” when used with respect to “ameliorating one ormore symptoms of eye disease” refers to a reduction, prevention, orelimination of one or more symptoms characteristic of eye disease and/orassociated pathologies. Such a reduction includes, but is not limited toa reduction or elimination of oxidized phospholipids, accumulation ofextracellular lipids in Bruch's membranes, accumulation of lipid richdebris in Bruch's membranes, vision loss, formation of choriocapillaris,thickening of the Bruch's membrane, accumulation of neutral lipids inthe Bruch's membrane, formation of a diffusion barrier between theretinal pigment epithelium, deposition of debris (basal linear depositsand drusen) between the basal membrane of the RPE and the innercollagenous layer, accumulation of lipofuscin in the RPE cells, RPEatrophy, photoreceptor degeneration, choroidal neovascularization,trapped fluid accumulation in the retina and retinal pigment epithelialcells, elevated intraocular pressure, as well as leakage, bleeding,scarring of the eye, and the like.

The term “enantiomeric amino acids” refers to amino acids that can existin at least two forms that are nonsuperimposable mirror images of eachother. Most amino acids (except glycine) are enantiomeric and exist in aso-called L-form (L amino acid) or D-form (D amino acid). Most naturallyoccurring amino acids are “L” amino acids. The terms “D amino acid” and“L amino acid” are used to refer to absolute configuration of the aminoacid, rather than a particular direction of rotation of plane-polarizedlight. The usage herein is consistent with standard usage by those ofskill in the art. Amino acids are designated herein using standard1-letter or three-letter codes, e.g. as designated in Standard ST.25 inthe Handbook On Industrial Property Information and Documentation.

The term “protecting group” refers to a chemical group that, whenattached to a functional group in an amino acid (e.g. a side chain, analpha amino group, an alpha carboxyl group, etc.) blocks or masks theproperties of that functional group. Preferred amino-terminal protectinggroups include, but are not limited to acetyl, or amino groups. Otheramino-terminal protecting groups include, but are not limited to alkylchains as in fatty acids, propeonyl, formyl and others. Preferredcarboxyl terminal protecting groups include, but are not limited togroups that form amides or esters.

The phrase “protect a phospholipid from oxidation by an oxidizing agent”refers to the ability of a compound to reduce the rate of oxidation of aphospholipid (or the amount of oxidized phospholipid produced) when thatphospholipid is contacted with an oxidizing agent (e.g. hydrogenperoxide, 13-(S)-HPODE, 15-(S)-HPETE, HPODE, HPETE, HODE, HETE, etc.).

The terms “low density lipoprotein” or “LDL” is defined in accordancewith common usage of those of skill in the art. Generally, LDL refers tothe lipid-protein complex which when isolated by ultracentrifugation isfound in the density range d=1.019 to d=1.063.

The terms “high density lipoprotein” or “HDL” is defined in accordancewith common usage of those of skill in the art. Generally “HDL” refersto a lipid-protein complex which when isolated by ultracentrifugation isfound in the density range of d=1.063 to d=1.21.

The term “Group I HDL” refers to a high density lipoprotein orcomponents thereof (e.g. apo A-I, paraoxonase, platelet activatingfactor acetylhydrolase, etc.) that reduce oxidized lipids (e.g. in lowdensity lipoproteins) or that protect oxidized lipids from oxidation byoxidizing agents.

The term “Group II HDL” refers to an HDL that offers reduced activity orno activity in protecting lipids from oxidation or in repairing (e.g.reducing) oxidized lipids.

The term “HDL component” refers to a component (e.g. molecules) thatcomprises a high density lipoprotein (HDL). Assays for HDL that protectlipids from oxidation or that repair (e.g. reduce oxidized lipids) alsoinclude assays for components of HDL (e.g. apo A-I, paraoxonase,platelet activating factor acetylhydrolase, etc.) that display suchactivity.

The term “human apo A-I peptide” refers to a full-length human apo A-Ipeptide or to a fragment or domain thereof comprising a class Aamphipathic helix.

A “monocytic reaction” as used herein refers to monocyte activitycharacteristic of the “inflammatory response” associated withatherosclerotic plaque formation. The monocytic reaction ischaracterized by monocyte adhesion to cells of the vascular wall (e.g.cells of the vascular endothelium), and/or chemotaxis into thesubendothelial space, and/or differentiation of monocytes intomacrophages.

The term “absence of change” when referring to the amount of oxidizedphospholipid refers to the lack of a detectable change, more preferablythe lack of a statistically significant change (e.g. at least at the85%, preferably at least at the 90%, more preferably at least at the95%, and most preferably at least at the 98% or 99% confidence level).The absence of a detectable change can also refer to assays in whichoxidized phospholipid level changes, but not as much as in the absenceof the protein(s) described herein or with reference to other positiveor negative controls.

The following abbreviations may be used herein: PAPC:L-α-1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine; POVPC:1-palmitoyl-2-(5-oxovaleryl)-sn-glycero-3-phosphocholine; PGPC:1-palmitoyl-2-glutaryl-sn-glycero-3-phosphocholine; PEIPC:1-palmitoyl-2-(5,6-epoxyisoprostane E₂)-sn-glycero-3-phosphocholine;ChC18:2: cholesteryl linoleate; ChC18:2-OOH: cholesteryl linoleatehydroperoxide; DMPC: 1,2-ditetradecanoyl-rac-glycerol-3-phosphocholine;PON: paraoxonase; HPF: Standardized high power field; PAPC:L-α-1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine;BL/6:C57BL/6J; C3H:C3H/HeJ.

The term “conservative substitution” is used in reference to proteins orpeptides to reflect amino acid substitutions that do not substantiallyalter the activity (specificity (e.g. for lipoproteins)) or bindingaffinity (e.g. for lipids or lipoproteins)) of the molecule. Typicallyconservative amino acid substitutions involve substitution one aminoacid for another amino acid with similar chemical properties (e.g.charge or hydrophobicity). The following six groups each contain aminoacids that are typical conservative substitutions for one another: 1)Alanine (A), Serine (S), Threonine (T); 2) Aspartic acid (D), Glutamicacid (E); 3) Asparagine (N), Glutamine (Q); 4) Arginine (R), Lysine (K);5) Isoleucine (I), Leucine (L), Methionine (M), Valine (V); and 6)Phenylalanine (F), Tyrosine (Y), Tryptophan (W).

The terms “identical” or percent “identity,” in the context of two ormore nucleic acids or polypeptide sequences, refer to two or moresequences or subsequences that are the same or have a specifiedpercentage of amino acid residues or nucleotides that are the same, whencompared and aligned for maximum correspondence, as measured using oneof the following sequence comparison algorithms or by visual inspection.With respect to the peptides of this invention sequence identity isdetermined over the full length of the peptide.

For sequence comparison, typically one sequence acts as a referencesequence, to which test sequences are compared. When using a sequencecomparison algorithm, test and reference sequences are input into acomputer, subsequence coordinates are designated, if necessary, andsequence algorithm program parameters are designated. The sequencecomparison algorithm then calculates the percent sequence identity forthe test sequence(s) relative to the reference sequence, based on thedesignated program parameters.

Optimal alignment of sequences for comparison can be conducted, e.g., bythe local homology algorithm of Smith & Waterman, Adv. Appl. Math. 2:482(1981), by the homology alignment algorithm of Needleman & Wunsch, J.Mol. Biol. 48:443 (1970), by the search for similarity method of Pearson& Lipman (1988) Proc. Natl. Acad. Sci. USA 85:2444, by computerizedimplementations of these algorithms (GAP, BESTFIT, FASTA, and TFASTA inthe Wisconsin Genetics Software Package, Genetics Computer Group, 575Science Dr., Madison, Wis.), or by visual inspection (see generallyAusubel et al., supra).

One example of a useful algorithm is PILEUP. PILEUP creates a multiplesequence alignment from a group of related sequences using progressive,pairwise alignments to show relationship and percent sequence identity.It also plots a tree or dendogram showing the clustering relationshipsused to create the alignment. PILEUP uses a simplification of theprogressive alignment method of Feng & Doolittle (1987) J. Mol. Evol.35:351-360. The method used is similar to the method described byHiggins & Sharp (1989) CABIOS 5: 151-153. The program can align up to300 sequences, each of a maximum length of 5,000 nucleotides or aminoacids. The multiple alignment procedure begins with the pairwisealignment of the two most similar sequences, producing a cluster of twoaligned sequences. This cluster is then aligned to the next most relatedsequence or cluster of aligned sequences. Two clusters of sequences arealigned by a simple extension of the pairwise alignment of twoindividual sequences. The final alignment is achieved by a series ofprogressive, pairwise alignments. The program is run by designatingspecific sequences and their amino acid or nucleotide coordinates forregions of sequence comparison and by designating the programparameters. For example, a reference sequence can be compared to othertest sequences to determine the percent sequence identity relationshipusing the following parameters: default gap weight (3.00), default gaplength weight (0.10), and weighted end gaps.

Another example of algorithm that is suitable for determining percentsequence identity and sequence similarity is the BLAST algorithm, whichis described in Altschul et al. (1990) J. Mol. Biol. 215: 403-410.Software for performing BLAST analyses is publicly available through theNational Center for Biotechnology Information(http://www.ncbi.nlm.nih.gov/). This algorithm involves firstidentifying high scoring sequence pairs (HSPs) by identifying shortwords of length W in the query sequence, which either match or satisfysome positive-valued threshold score T when aligned with a word of thesame length in a database sequence. T is referred to as the neighborhoodword score threshold (Altschul et al, supra). These initial neighborhoodword hits act as seeds for initiating searches to find longer HSPscontaining them. The word hits are then extended in both directionsalong each sequence for as far as the cumulative alignment score can beincreased. Cumulative scores are calculated using, for nucleotidesequences, the parameters M (reward score for a pair of matchingresidues; always >0) and N (penalty score for mismatching residues;always <0). For amino acid sequences, a scoring matrix is used tocalculate the cumulative score. Extension of the word hits in eachdirection are halted when: the cumulative alignment score falls off bythe quantity X from its maximum achieved value; the cumulative scoregoes to zero or below, due to the accumulation of one or morenegative-scoring residue alignments; or the end of either sequence isreached. The BLAST algorithm parameters W, T, and X determine thesensitivity and speed of the alignment. The BLASTN program (fornucleotide sequences) uses as defaults a wordlength (W) of 11, anexpectation (E) of 10, M=5, N=−4, and a comparison of both strands. Foramino acid sequences, the BLASTP program uses as defaults a wordlength(W) of 3, an expectation (E) of 10, and the BLOSUM62 scoring matrix (seeHenikoff & Henikoff (1989) Proc. Natl. Acad. Sci. USA 89:10915).

In addition to calculating percent sequence identity, the BLASTalgorithm also performs a statistical analysis of the similarity betweentwo sequences (see, e.g., Karlin & Altschul (1993) Proc. Natl. Acad.Sci. USA, 90: 5873-5787). One measure of similarity provided by theBLAST algorithm is the smallest sum probability (P(N)), which providesan indication of the probability by which a match between two nucleotideor amino acid sequences would occur by chance. For example, a nucleicacid is considered similar to a reference sequence if the smallest sumprobability in a comparison of the test nucleic acid to the referencenucleic acid is less than about 0.1, more preferably less than about0.01, and most preferably less than about 0.001.

The phrase “in conjunction with” when used in reference to the use ofone or more drugs in conjunction with one or more active agentsdescribed herein indicates that the drug(s) and the active agent(s) areadministered so that there is at least some chronological overlap intheir physiological activity on the organism. Thus the drug(s) andactive agent(s) can be administered simultaneously and/or sequentially.In sequential administration there may even be some substantial delay(e.g., minutes or even hours or days) before administration of thesecond moiety as long as the first administered drug/agent has exertedsome physiological alteration on the organism when the secondadministered agent is administered or becomes active in the organism.

The phrases “adjacent to each other in a helical wheel diagram of apeptide” or “contiguous in a helical wheel diagram of a peptide” whenreferring to residues in a helical peptide indicates that in the helicalwheel representation the residues appear adjacent or contiguous eventhough they may not be adjacent or contiguous in the linear peptide.Thus, for example, the residues “A, E, K, W, K, and F” are contiguous inthe helical wheel diagrams shown in FIG. 15 even though these residuesare not contiguous in the linear peptide.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows a comparison of the effect of D4F (Navab, et al. (2002)Circulation, 105: 290-292) and apo-J peptide 336 made from D amino acids(D-J336*) on the prevention of LDL-induced monocyte chemotactic activityin vitro in a co-incubation experiment. The data are mean±SD of thenumber of migrated monocytes in nine high power fields in quadruplecultures. (D-J336=Ac-LLEQLNEQFNWVSRLANLTQGE-NH₂, SEQ ID NO:1011).

FIG. 2 illustrates the prevention of LDL-induced monocyte chemotacticactivity by pre-treatment of artery wall cells with D-J336 as comparedto D-4F. The data are mean±SD of the number of migrated monocytes innine high power fields in quadruple cultures.

FIG. 3 illustrates the effect of apo J peptide mimetics on HDLprotective capacity in LDL receptor null mice. The values are themean±SD of the number of migrated monocytes in 9 high power fields fromeach of quadruple assay wells.

FIG. 4 illustrates protection against LDL-induced monocyte chemotacticactivity by HDL from apo E null mice given oral peptides. The values arethe mean±SD of the number of migrated monocytes in 9 high power fieldsfrom each of quadruple assay wells. Asterisks indicate significantdifference (p<0.05) as compared to No Peptide mHDL.

FIG. 5 illustrates the effect of oral apo A-1 peptide mimetic and apoJpeptide on LDL susceptibility to oxidation. The values are the mean±SDof the number of migrated monocytes in 9 high power fields from each ofquadruple assay wells. Asterisks indicate significant difference(p<0.05) as compared to No Peptide LDL.

FIG. 6 illustrates the effect of oral apoA-1 peptide mimetic and apoJpeptide on HDL protective capacity. The values are the mean±SD of thenumber of migrated monocytes in 9 high power fields from each ofquadruple assay wells. Asterisks indicate significant difference(p<0.05) as compared to No Peptide mHDL.

FIG. 7 illustrates the effect of oral apoA-1 peptide mimetic and apoJpeptide on plasma paraoxonase activity. The values are the mean±SD ofreadings from quadruple plasma aliquots. Asterisks indicate significantdifferences (p<0.05) as compared to No Peptide control plasma.

FIG. 8 shows the effect of oral G* peptides on HDL protective capacityin apoE−/− mice. The values are the mean±SD of readings from quadrupleplasma aliquots. Asterisks indicate significant differences (p<0.05) ascompared to no peptide control plasma.

FIG. 9 shows the effect of Oral G* peptide, 146-156, on HDL protectivecapacity in ApoE−/− mice.

FIGS. 10A through 10C illustrate helical wheel diagrams of certainpeptides of this invention. FIG. 10A: V²W³A⁵F^(10,17)-D-4F (SEQ ID NO.1168); FIG. 10B: W³-D-4F (SEQ ID NO. 1132); FIG. 10C: V²W³F¹⁰-D-4F (SEQID NO. 1169).

FIG. 11A standard human LDL (LDL) was added to human artery wallcocultures without (No Addition) or with human HDL (+Control HDL) orwith mouse HDL from apoE null mice given Chow overnight (+Chow HDL), orgiven D-4F in the chow overnight (+D4F HDL) or given G5-D-4F in the chowovernight (+G5 HDL), or given G5,10-D-4F in the chow overnight (+5-10HDL), or given G5,11-D-4F in the chow overnight (+5-11 HDL) and theresulting monocyte chemotactic activity determined as previouslydescribed (Navab et al. (2002) Circulation, 105: 290-292).

FIG. 12 shows that peptides of this invention are effective inmitigating symptoms of diabetes (e.g., blood glucose). Obese Zucker rats26 weeks of age were bled and then treated with daily intraperitonealinjections of D-4F (5.0 mg/kg/day). After 10 days the rats were bledagain plasma glucose and lipid hydroperoxides (LOOH) were determined.*p=0.027; **p=0.0017.

FIG. 13. Sixteen week old Obese Zucker Rats were injected with D-4F (5mg/kg/daily) for 1 week at which time they underwent balloon injury ofthe common carotid artery. Two weeks later the rats were sacrificed andthe intimal media ratio determined.

FIG. 14 demonstrates that the product of the solution phase synthesisscheme is very biologically active in producing HDL and pre-beta HDLthat inhibit LDL-induced monocyte chemotaxis in apo E null mice. ApoEnull mice were fed 5 micrograms of the D-4F synthesized as describedabove (Frgmnt) or the mice were given the same amount of mouse chowwithout D-4F (Chow). Twelve hours after the feeding was started, themice were bled and their plasma was fractionated on FPLC. LDL (100micrograms LDL-cholesterol) was added to cocultures of human artery wallcells alone (LDL) or with a control human HDL (Control HDL) or with HDL(50 micrograms HDL-cholesterol) or post-HDL (pHDL; prebeta HDL) frommice that did (Frgmnt) or did not (Chow) receive the D-4F and themonocyte chemotactic activity produced was determined

FIG. 15 illustrates a helical wheel representation of 4F and reverse(retro) 4F. Reverse-4F is a mirror image of 4F with the relativepositions of the amino acids to each other and to the hydrophilic andhydrophobic faces being identical.

FIG. 16 shows a comparison of the HDL inflammatory index of D-4F versusreverse D-4F.

FIG. 17A (1A from Rudolf Summary) shows electron microscopy of untreatedeyes (controls) at a magnification of 5,000×; Bruch's membrane (BrM,arrow heads) structure loosen up with many translucent lipid vacuoles,these degenerative changes can be observed in both control eyes in atleast ¾ of the entire Bruch's membrane. In every other RPE cell biglipid vacuoles (asterisks) are found, a sign of stress and beginningdegeneration. PR: photo receptors; CC: choriocapillaris: EC: vascularendothelial cell of the choriocapillaris.

FIG. 17B (1B from Rudolf Summary) shows electron microscopy of untreatedeyes (controls) at a magnification of 20,000×; Bruch's membrane (BrM,arrow heads) shows a significant compromised morphology without theregular and uniform five layer arrangement. The membrane is slightlythickened with multiple translucent lipid vacuoles. A few are markedwith asterisks. EC: vascular endothelial cell of choriocapillaris.

FIG. 18A (2A from Rudolf Summary) shows electron microscopy of treatedeyes (Apolipoprotein mimetic peptide L-4F) at a magnification of 5,000×;regular and uniform Bruch's membrane structure (BrM, black arrow heads)which resembles Bruch's membrane in healthy wild type mice (Dithmar etal. (2000) Invest Opthalmol Vis Sci 41:2035-42); only occasionally alipid vacuole left (white arrow head); this effect was found in theentire Bruch's membrane; none of the RPE cells showed lipid droplets.

FIG. 18B (2B from Rudolf Summary) shows electron microscopy of a treatedeye (Apolipoprotein mimetic peptide L-4F) at a magnification of 20,000×;regular and uniform Bruch's membrane (BrM, black arrow heads) notranslucent lipid vacuoles shown.

DETAILED DESCRIPTION I. Methods of Treatment

The active agents (e.g. peptides, small organic molecules, amino acidpairs, etc.) described herein are effective for mitigating one or moresymptoms and/or reducing the rate of onset and/or severity of one ormore indications described herein. In particular, the active agents(e.g. peptides, small organic molecules, amino acid pairs, etc.)described herein are effective for mitigating one or more symptoms ofatherosclerosis and/or eye disease. Without being bound to a particulartheory, it is believed that the peptides bind the “seeding molecules”required for the formation of pro-inflammatory oxidized phospholipidssuch as Ox-PAPC, POVPC, PGPC, and PEIPC.

In addition, since many inflammatory conditions and/or other pathologiesare mediated at least in part by oxidized lipids, we believe that thepeptides of this invention are effective in ameliorating conditions thatare characterized by the formation of biologically active oxidizedlipids. In addition, there are a number of other conditions for whichthe active agents described herein appear to be efficacious.

A number of pathologies for which the active agents described hereinappear to be a palliative and/or a preventative are described below.

A) Atherosclerosis and Associated Pathologies.

We discovered that normal HDL inhibits three steps in the formation ofmildly oxidized LDL. In particular, we demonstrated that treating humanLDL in vitro with apo A-I or an apo A-I mimetic peptide (37 pA) removedseeding molecules from the LDL that included HPODE and HPETE. Theseseeding molecules were required for cocultures of human artery wallcells to be able to oxidize LDL and for the LDL to induce the arterywall cells to produce monocyte chemotactic activity. We alsodemonstrated that after injection of apo A-I into mice or infusion intohumans, the LDL isolated from the mice or human volunteers afterinjection/infusion of apo A-I was resistant to oxidation by human arterywall cells and did not induce monocyte chemotactic activity in theartery wall cell cocultures.

The protective function of various active agents of this invention isillustrated in the parent applications (Ser. No. 09/645,454, filed Aug.24, 2000, Ser. No. 09/896,841, filed Jun. 29, 2001, and WO 02/15923(PCT/US01/26497), filed Jun. 29, 2001, see, e.g., FIGS. 1-5 in WO02/15923. FIG. 1, panels A, B, C, and D in WO 02/15923 show theassociation of 14C-D-5F with blood components in an ApoE null mouse). Itis also demonstrated that HDL from mice that were fed an atherogenicdiet and injected with PBS failed to inhibit the oxidation of human LDLand failed to inhibit LDL-induced monocyte chemotactic activity in humanartery wall cocultures. In contrast, HDL from mice fed an atherogenicdiet and injected daily with peptides described herein was as effectivein inhibiting human LDL oxidation and preventing LDL-induced monocytechemotactic activity in the cocultures as was normal human HDL (FIGS. 2Aand 2B in WO 02/15923). In addition, LDL taken from mice fed theatherogenic diet and injected daily with PBS was more readily oxidizedand more readily induced monocyte chemotactic activity than LDL takenfrom mice fed the same diet but injected with 20.mu.g daily of peptide5F. The D peptide did not appear to be immunogenic (FIG. 4 in WO02/15923).

The in vitro responses of human artery wall cells to HDL and LDL frommice fed the atherogenic diet and injected with a peptide according tothis invention are consistent with the protective action shown by suchpeptides in vivo. Despite, similar levels of total cholesterol,LDL-cholesterol, IDL+VLDL-cholesterol, and lower HDL-cholesterol as apercent of total cholesterol, the animals fed the atherogenic diet andinjected with the peptide had significantly lower lesion scores (FIG. 5in WO 02/15923). The peptides of this invention thus preventedprogression of atherosclerotic lesions in mice fed an atherogenic diet.

Thus, in one embodiment, this invention provides methods forameliorating and/or preventing one or more symptoms of atherosclerosisby administering one or more of the active agents described herein.

It is also noted that c-reactive protein, a marker for inflammation, iselevated in congestive heart failure. Also, in congestive heart failurethere is an accumulation of reactive oxygen species and vasomotionabnormalities. Because of their effects in preventing/reducing theformation of various oxidized species and/or because of their effect inimproving vasoreactivity and/or arteriole function (see below) theactive agents described herein will be effective in treating congestiveheart failure.

B) Arteriole/Vascular Indications.

Vessels smaller than even the smallest arteries (i.e., arterioles)thicken, become dysfunctional and cause end organ damage to tissues asdiverse as the brain and the kidney. It is believed the active agentsdescribed herein can function to improve areteriole structure andfunction and/or to slow the rate and/or severity of arterioledysfunction. Without being bound to a particular theory, it is believedthat arteriole dysfunction is a causal factor in various brain andkidney disorders. Use of the agents described herein thus provides amethod to improve the structure and function of arterioles and preservethe function of end organs such as the brain and kidney.

Thus, for example, administration of one or more of the active agentsdescribed herein is expected to reduce one or more symptoms or to slowthe onset or severity of arteriolar disease associated with aging,and/or Alzheimer's disease, and/or Parkinson's disease, and/or withmulti-infarct dementia, and/or subarachnoid hemorrhage, and the like.Similarly, administration of one or more agents described herein isexpected to mitigate one or more symptoms and/or to slow the onsetand/or severity of chronic kidney disease, and/or hypertension.

Similarly, the agents described herein appear to improve vasoreactivity.Because of the improvement of vasoreactivity and/or arteriole function,the agents described herein are suitable for the treatment of peripheralvascular disease, erectile dysfunction, and the like.

C) Pulmonary Indications.

The agents described herein are also suitable for treatment of a varietyof pulmonary indications. These include, but are not limited to chronicobstructive pulmonary disease (COPD), emphysema, pulmonary disease,asthma, idiopathic pulmonary fibrosis, and the like.

D) Mitigation of a Symptom or Condition Associated with CoronaryCalcification and Osteoporosis.

Vascular calcification and osteoporosis often co-exist in the samesubjects (Ouchi et al. (1993) Ann NY Acad. Sci., 676: 297-307; Boukhrisand Becker (1972) JAMA, 219: 1307-1131; Banks et al. (1994) Eur J ClinInvest., 24: 813-817; Laroche et al. (1994) Clin Rheumatol., 13:611-614; Broulik and Kapitola (1993) Endocr Regul., 27: 57-60; Frye etal. (1992) Bone Mine., 19: 185-194; Barengolts et al. (1998) CalcifTissue Int., 62: 209-213; Burnett and Vasikaran (2002) Ann ClinBiochem., 39: 203-210. Parhami et al. (1997) Arterioscl Thromb VascBiol., 17: 680-687, demonstrated that mildly oxidized LDL (MM-LDL) andthe biologically active lipids in MM-LDL [i.e. oxidized1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphorylcholine) (Ox-PAPC)],as well as the isoprostane, 8-iso prostaglandin F₂, but not theunoxidized phospholipid (PAPC) or isoprostane 8-iso progstaglandin F₂αinduced alkaline phosphatase activity and osteoblastic differentiationof calcifying vascular cells (CVCs) in vitro, but inhibited thedifferentiation of MC3T3-E1 bone cells.

The osteon resembles the artery wall in that the osteon is centered onan endothelial cell-lined lumen surrounded by a subendothelial spacecontaining matrix and fibroblast-like cells, which is in turn surroundedby preosteoblasts and osteoblasts occupying a position analogous tosmooth muscle cells in the artery wall (Id.). Trabecular boneosteoblasts also interface with bone marrow subendothelial spaces (Id.).Parhami et al. postulated that lipoproteins could cross the endotheliumof bone arteries and be deposited in the subendothelial space where theycould undergo oxidation as in coronary arteries (Id.). Based on their invitro data they predicted that LDL oxidation in the subendothelial spaceof bone arteries and in bone marrow would lead to reduced osteoblasticdifferentiation and mineralization which would contribute toosteoporosis (Id.). Their hypothesis further predicted that LDL levelswould be positively correlated with osteoporosis as they are withcoronary calcification (Pohle et al. (2001) Circulation, 104:1927-1932), but HDL levels would be negatively correlated withosteoporosis (Parhami et al. (1997) Arterioscl Thromb Vase Biol., 17:680-687).

In vitro, the osteoblastic differentiation of the marrow stromal cellline M2-10B4 was inhibited by MM-LDL but not native LDL (Parhami et al.(1999) J Bone Miner Res., 14: 2067-2078). When marrow stromal cells fromatherosclerosis susceptible C57BU6 (BL6) mice fed a low fat chow dietwere cultured there was robust osteogenic differentiation (Id.). Incontrast, when the marrow stromal cells taken from the mice after a highfat, atherogenic diet were cultured they did not undergo osteogenicdifferentiation (Id.). This observation is particularly important sinceit provides a possible explanation for the decreased osteogenicpotential of marrow stromal cells in the development of osteoporosis(Nuttall and Gimble (2000) Bone, 27: 177-184). In vivo the decrease inosteogenic potential is accompanied by an increase in adipogenesis inosteoporotic bone (Id.).

It was found that adding D-4F to the drinking water of apoE null micefor 6 weeks dramatically increased trabecular bone mineral density andit is believed that the other active agents of this invention will actsimilarly.

Our data indicate that osteoporosis can be regarded as an“atherosclerosis of bone”. It appears to be a result of the action ofoxidized lipids. HDL destroys these oxidized lipids and promotesosteoblastic differentiation. Our data indicate that administeringactive agent (s) of this invention to a mammal (e.g., in the drinkingwater of apoE null mice) dramatically increases trabecular bone in justa matter of weeks.

This indicates that the active agents, described herein are useful formitigation one or more symptoms of osteoporosis (e.g., for inhibitingdecalcification) or for inducing recalcification of osteoporotic bone.The active agents are also useful as prophylactics to prevent the onsetof symptom(s) of osteoporosis in a mammal (e.g., a patient at risk forosteoporosis).

We believe similar mechanisms are a cause of coronary calcification,e.g., calcific aortic stenosis. Thus, in certain embodiments, thisinvention contemplates the use of the active agents described herein toinhibit or prevent a symptom of a disease such as coronarycalcification, calcific aortic stenosis, osteoporosis, and the like.

E) Inflammatory and Autoimmune Indications.

Chronic inflammatory and/or autoimmune conditions are also characterizedby the formation of a number of reactive oxygen species and are amenableto treatment using one or more of the active agents described herein.Thus, without being bound to a particular theory, we also believe theactive agents described herein are useful, prophylactically ortherapeutically, to mitigate the onset and/or more or more symptoms of avariety of other conditions including, but not limited to rheumatoidarthritis, lupus erythematous, polyarteritis nodosa, polymyalgiarheumatica, scleroderma, multiple sclerosis, and the like.

In certain embodiments, the active agents are useful in mitigating oneor more symptoms caused by, or associated with, an inflammatory responsein these conditions.

Also, in certain embodiments, the active agents are useful in mitigatingone or more symptoms caused by or associated with an inflammatoryresponse associated with AIDS.

F) Infections/Trauma/Transplants.

We have observed that a consequence of influenza infection and otherinfections is the diminution in paraoxonase and platelet activatingacetylhydrolase activity in the HDL. Without being bound by a particulartheory, we believe that, as a result of the loss of these HDL enzymaticactivities and also as a result of the association of pro-oxidantproteins with HDL during the acute phase response, HDL is no longer ableto prevent LDL oxidation and is no longer able to prevent theLDL-induced production of monocyte chemotactic activity by endothelialcells.

We observed that in a subject injected with very low dosages of certainagents of this invention (e.g., 20 micrograms for mice) daily afterinfection with the influenza A virus paraoxonase levels did not fall andthe biologically active oxidized phospholipids were not generated beyondbackground. This indicates that 4F, D4F (and/or other agents of thisinvention) can be administered (e.g. orally or by injection) to patients(including, for example with known coronary artery disease duringinfluenza infection or other events that can generate an acute phaseinflammatory response, e.g. due to viral infection, bacterial infection,trauma, transplant, various autoimmune conditions, etc.) and thus we canprevent by this short term treatment the increased incidence of heartattack and stroke associated with pathologies that generate suchinflammatory states.

In addition, by restoring and/or maintaining paroxonase levels and/ormonocyte activity, the agent(s) of this invention are useful in thetreatment of infection (e.g., viral infection, bacterial infection,fungal infection) and/or the inflammatory pathologies associated withinfection (e.g. meningitis) and/or trauma.

In certain embodiments, because of the combined anti-inflammatoryactivity and anti-infective activity, the agents described herein arealso useful in the treatment of a wound or other trauma, mitigatingadverse effects associated with organ or tissue transplant, and/or organor tissue transplant rejection, and/or implanted prostheses, and/ortransplant atherosclerosis, and/or biofilm formation. In addition, webelieve that L-4F, D-4F, and/or other agents described herein are alsouseful in mitigating the effects of spinal cord injuries.

G) Diabetes and Associated Conditions.

Various active agents described herein have also been observed to showefficacy in reducing and/or preventing one or more symptoms associatedwith diabetes. Thus, in various embodiments, this invention providesmethods of treating (therapeutically and/or prophylactically) diabetesand/or associated pathologies (e.g., Type I diabetes, Type II diabetes,juvenile onset diabetes, diabetic nephropathy, nephropathy, diabeticneuropathy, diabetic retinopathy, and the like.

H) Cancer.

NFκB is a transcription factor that is normally activated in response toproinflammatory cytokines and that regulates the expression of more than200 genes. Many tumor cell lines show constitutive activation of NFκBsignaling. Various studies of mouse models of intestinal, and mammarytumors conclude that activation of the NFκB pathway enhances tumordevelopment and may act primarily in the late stages of tumorigenesis(see, e.g., (2004) Cell 118: 285; (2004) J. Clin. Invest., 114: 569).Inhibition of NFκB signaling suppressed tumor development. Without beingbound to a particular theory, mechanisms for this suppression arebelieved to include an increase in tumor cell apoptosis, reducedexpression of tumor cell growth factors supplied by surrounding stromalcells, and/or abrogation of a tumor cell dedifferentiation program thatis critical for tumor invasion/metastasis.

Without being bound by a particular theory, it is believed theadministration of one or more active agents described herein willinhibit expression and/or secretion, and/or activity of NFκB. Thus, incertain embodiments, this invention provides methods of treating apathology characterized by elevated NFκB by administering one or moreactive agents described herein. Thus, In various embodiments thisinvention contemplates inhibiting NFκB activation associated with cancerby administering one or more active agents described herein, optionallyin combination with appropriate cancer therapeutics.

I) Biochemical Activity.

The active agent(s) described herein have been shown to exhibit a numberof specific biological activities. Thus, for example, they increase hemeoxygenase 1, they increase extracellular superoxide dismutase, theyreduce or prevent the association of myeloperoxidase with apoA-I, theyreduce or prevent the nitrosylation of tyrosine in apoA-I, they renderHDL Anti-inflammatory or more anti-inflammatory, and they increase theformation cycling of pre-βHDL, they promote reverse cholesteroltransport, in particular, reverse cholesterol transport frommacrophages, and they synergize the activity of statins. The activeagents described herein can thus be administered to a mammal to promoteany of these activities, e.g. to treat a condition/pathology whoseseverity, and/or likelihood of onset is reduced by one or more of theseactivities.

J) Mitigation of a Symptom of Atherosclerosis Associated with an AcuteInflammatory Response.

The active agents, of this invention are also useful in a number ofcontexts. For example, we have observed that cardiovascularcomplications (e.g., atherosclerosis, stroke, etc.) frequently accompanyor follow the onset of an acute phase inflammatory response, e.g., suchas that associated with a recurrent inflammatory disease, a viralinfection (e.g., influenza), a bacterial infection, a fungal infection,an organ transplant, a wound or other trauma, and so forth.

Thus, in certain embodiments, this invention contemplates administeringone or more of the active agents described herein to a subject at riskfor, or incurring, an acute inflammatory response and/or at risk for orincurring a symptom of atherosclerosis and/or an associated pathology(e.g., stroke).

Thus, for example, a person having or at risk for coronary disease mayprophylactically be administered a one or more active agents of thisinvention during flu season. A person (or animal) subject to a recurrentinflammatory condition, e.g., rheumatoid arthritis, various autoimmunediseases, etc., can be treated with a one or more agents describedherein to mitigate or prevent the development of atherosclerosis orstroke. A person (or animal) subject to trauma, e.g., acute injury,tissue transplant, etc. can be treated with a polypeptide of thisinvention to mitigate the development of atherosclerosis or stroke.

In certain instances such methods will entail a diagnosis of theoccurrence or risk of an acute inflammatory response. The acuteinflammatory response typically involves alterations in metabolism andgene regulation in the liver. It is a dynamic homeostatic process thatinvolves all of the major systems of the body, in addition to theimmune, cardiovascular and central nervous system. Normally, the acutephase response lasts only a few days; however, in cases of chronic orrecurring inflammation, an aberrant continuation of some aspects of theacute phase response may contribute to the underlying tissue damage thataccompanies the disease, and may also lead to further complications, forexample cardiovascular diseases or protein deposition diseases such asamyloidosis.

An important aspect of the acute phase response is the radically alteredbiosynthetic profile of the liver. Under normal circumstances, the liversynthesizes a characteristic range of plasma proteins at steady stateconcentrations. Many of these proteins have important functions andhigher plasma levels of these acute phase reactants (APRs) or acutephase proteins (APPs) are required during the acute phase responsefollowing an inflammatory stimulus. Although most APRs are synthesizedby hepatocytes, some are produced by other cell types, includingmonocytes, endothelial cells, fibroblasts and adipocytes. Most APRs areinduced between 50% and several-fold over normal levels. In contrast,the major APRs can increase to 1000-fold over normal levels. This groupincludes serum amyloid A (SAA) and either C-reactive protein (CRP) inhumans or its homologue in mice, serum amyloid P component (SAP).So-called negative APRs are decreased in plasma concentration during theacute phase response to allow an increase in the capacity of the liverto synthesize the induced APRs.

In certain embodiments, the acute phase response, or risk therefore isevaluated by measuring one or more APPs. Measuring such markers is wellknown to those of skill in the art, and commercial companies exist thatprovide such measurement (e.g., AGP measured by Cardiotech Services,Louisville, Ky.).

K) Eye Disease

Also disclosed are methods for ameliorating and/or preventing one ormore symptoms of eye disease by administering one or more of the activeagents described herein. As described above, the “eye disease” includes,but is not limited to, age related maculopathy (ARM), age relatedmacular degeneration (AMD) including both the dry and wet forms of agerelated macular degeneration, glaucoma, ocular hypertension, macularedema, retinal pigment epithelium detachments, coats disease, uveitis,sicca syndrome, hereditary diseases associated with increasedextra-/intracellular lipid storage/accumulation, juvenile maculardegeneration as well as all risk factors for each mentioned disease.

For example the active agents disclosed herein can mobilize and removeaccumulated intra-/extracellular lipid deposits in all eye structures.The active agents can also structurally remodel essential transportpassages and supply structures and/or they have the highest affinity tooxidized lipids, which removal of such oxidized lipids can causes ananti-inflammatory effect.

The active agents can be used prophylactically in both the treatment andprevention of eye diseases if risk factors are present. Risk factors foreye disease are described elsewhere herein.

1) Macular Degeneration.

Age-related macular degeneration sometimes begins with characteristicyellow deposits in the macula (central area of the retina which providesdetailed central vision) called drusen between the retinal pigmentepithelium and the underlying choroid. Most people with these earlychanges (sometimes referred to as age-related maculopathy) have goodvision. People with drusen can go on to develop advanced maculardegeneration. The risk is considerably higher when the drusen are largeand numerous and associated with disturbance in the pigmented cell layerunder the macula. Recent research suggests that large and soft drusenare related to elevated cholesterol deposits and may respond tocholesterol lowering agents or the Rheo Procedure.

Advanced AMD, which is responsible for profound vision loss, has twoforms: dry and wet. Central geographic atrophy, the dry form of advancedAMD, results from atrophy to the retinal pigment epithelial layer belowthe retina, which causes vision loss through loss of photoreceptors(rods and cones) in the central part of the eye. While no treatment isavailable for this condition, vitamin supplements with high doses ofantioxidants, lutein and zeaxanthin, have been demonstrated by theNational Eye Institute and others to slow the progression of dry maculardegeneration and in some patients, improve visual acuity.

Neovascular or exudative AMD, the wet form of advanced AMD, causesvision loss due to abnormal blood vessel growth in thechoriocapillaries, through Bruch's membrane, ultimately leading to bloodand protein leakage below the macula. Bleeding, leaking, and scarringfrom these blood vessels eventually cause irreversible damage to thephotoreceptors and rapid vision loss if left untreated.

Until recently, no effective treatments were known for wet maculardegeneration. However, new drugs, called anti-angiogenics or anti-VEGF(anti-Vascular Endothelial Growth Factor) agents, when injected directlyinto the vitreous humor of the eye using a small, painless needle, cancause regression of the abnormal blood vessels and improvement ofvision. The injections frequently have to be repeated on a monthly orbi-monthly basis. Examples of these agents include Lucentis, Avastin andMacugen. Only Lucentis and Macugen are FDA approved as of April 2007.Macugen has been found to have only minimal benefits in neovascular AMDand is no longer used. Worldwide, Avastin has been used extensivelydespite its “off label” status. The cost of Lucentis is approximately$2000 US while the cost of Avastin is approximately $150.

2) AMD: Dry from: Geographic Atrophy,

The remodeling of Bruch's membrane provides an undisturbed passagebetween retinal pigment epithelium and choriocapillaris, which isessential for the health of the retina. The retinal pigment epitheliumstands with the choriocapillaris in a close relationship and they aredependent on each other. An uncompromised communication between thesestructures improves the blood supply for the outer retina by thechoriocapillaris and the retinal pigment epithelium layer integrity byimproved anchorage on Bruch's membrane via water soluble proteins.

3) AMD: Wet Form: Choroidal Neovascularization,

The same mechanism as for dry AMD applies. Due to the improvedenvironmental conditions retinal pigment epithelium cells also reducethe secretion of pro-angiogenic factors, which normally keeps aneovascularization active for a longer period. In combination withanti-angiogenic treatments (elsewhere herein) pro-angiogenic mechanismsare not just temporarily blocked but the secretion stimulus can belong-term reduced.

4) Glaucoma/Ocular Hypertension

One main characteristic of glaucoma/ocular hypertension is elevatedintraocular pressure (TOP). The treatment of the age-related “lipidwall” in Bruch's membrane increases the hydraulic conductivity alongBruch's membrane again and facilitates fluid transport from the vitreousto the choroid (vitreoretinal-choroidal outflow, uveoscleral outflow),which can normalize the IOP.

5) Macular Edema, Retinal Pigment Epithelium Detachments

Macular edema is characterized by trapped fluid accumulations in theretina and RPE detachments by fluid accumulations under the retinalpigment epithelium. The normal fluid transport is directed acrossBruch's membrane into the choriocapillaris. An improved transport acrossBruch's membrane after lipid clean up (“lipid wall”) and remodeling dueto ApoA-I/ApoE mimetic peptide treatment facilitates the natural fluidtransport and helps to resolve the sight-threatening macular edema andRPE detachment.

L) Mitigation of a Symptom of Macular Degeneration.

The active agents, of this invention are also useful in a number ofcontexts. For example, we have observed that eye disease (e.g., maculardegeneration, etc.) are frequently associated with drusen, basal lineardeposit, basal laminar deposit, lipid accumulation in Bruch's membrane,and/or positive genetic risk profiles, and so forth.

Thus, in certain embodiments, this invention contemplates administeringone or more of the active agents described herein to a subject at riskfor, or incurring, one or more of the symptoms and/or at risk for orincurring a symptom of an eye disease and/or an associated pathology(e.g., blindness).

Thus, for example, a person having or at risk for eye disease mayprophylactically be administered a one or more active agents of thisinvention during flu season. A person (or animal) subject to an eyedisease, e.g., macular degeneration, glaucoma, etc., can be treated witha one or more agents described herein to mitigate or prevent thedevelopment of eye disease. A person (or animal) subject to trauma,e.g., acute injury, tissue transplant, etc. can also be treated with apolypeptide of this invention to mitigate the development of eyedisease.

In certain instances such methods will entail a diagnosis of theoccurrence or risk of an eye disease. The eye disease typically involvesalterations in drusen, basal linear deposit, basal laminar deposit,lipid accumulation in and/or Bruch's membrane.

M) Other Indications.

In various embodiments it is contemplated that the active agentsdescribed herein are useful in the treatment (e.g. mitigation and/orprevention) of corneal ulcers, endothelial sloughing, Crohn's disease,acute and chronic dermatitis (including, but not limited to eczemaand/or psoriasis), macular degeneration, neuropathy, scleroderma, andulcerative colitis.

A summary of indications/conditions for which the active agents havebeen shown to be effective and/or are believed to be effective is shownin Table 1.

TABLE 1 Summary of conditions in which the active agents (e.g., D- 4F)have been shown to be or are believed to be effective.atherosclerosis/symptoms/consequences thereof plaque formation lesionformation myocardial infarction stroke congestive heart failure vascularfunction: arteriole function arteriolar disease associated with agingassociated with alzheimer's disease associated with chronic kidneydisease associated with hypertension associated with multi-infarctdementia associated with subarachnoid hemorrhage peripheral vasculardisease peripheral vascular disease pulmonary disease: chronicobstructive pulmonary disease (COPD) emphysema asthma idiopathicpulmonary fibrosis Pulmonary fibrosis adult respiratory distresssyndrome osteoporosis Paget's disease coronary calcification autoimmune:rheumatoid arthritis polyarteritis nodosa polymyalgia rheumatica lupuserythematosus multiple sclerosis Wegener's granulomatosis centralnervous system vasculitis (CNSV) Sjogren's syndrome Sclerodermapolymyositis AIDS inflammatory response infections: bacterial fungalviral parasitic influenza avian flu viral pneumonia endotoxic shocksyndrome sepsis sepsis syndrome (clinical syndrome where it appears thatthe patient is septic but no organisms are recovered from the blood)trauma/wound: organ transplant transplant atherosclerosis transplantrejection corneal ulcer chronic/non-healing wound ulcerative colitisreperfusion injury (prevent and/or treat) ischemic reperfusion injury(prevent and/or treat) spinal cord injuries (mitigating effects) cancersmyeloma/multiple myeloma ovarian cancer breast cancer colon cancer bonecancer osteoarthritis inflammatory bowel disease allergic rhinitiscachexia diabetes Alzheimer's disease implanted prosthesis biofilmformation Crohns' disease dermatitis, acute and chronic eczema psoriasiscontact dermatitis scleroderma diabetes and related conditions Type IDiabetes Type II Diabetes Juvenile Onset Diabetes Prevention of theonset of diabetes Diabetic Nephropathy Diabetic Neuropathy DiabeticRetinopathy erectile dysfunction macular degeneration multiple sclerosisnephropathy neuropathy Parkinson's Disease peripheral Vascular Diseasemeningitis Specific biological activities: increase Heme Oxygenase 1increase extracellular superoxide dismutase prevent endothelialsloughing prevent the association of myeloperoxidase with ApoA-I preventthe nitrosylation of tyrosine in ApoA-I render HDL anti-inflammatoryimprove vasoreactivity increase the formation of pre-beta HDL promotereverse cholesterol transport promote reverse cholesterol transport frommacrophages synergize the action of statins

It is noted that the conditions listed in Table 1 are intended to beillustrative and not limiting.

It is noted that the conditions listed in Table 1 are intended to beillustrative and not limiting.

N) Administration.

Typically the active agent(s) will be administered to a mammal (e.g., ahuman) in need thereof. Such a mammal will typically include a mammal(e.g. a human) having or at risk for one or more of the pathologiesdescribed herein.

The active agent(s) can be administered, as described herein, accordingto any of a number of standard methods including, but not limited toinjection, suppository, nasal spray, time-release implant, transdermalpatch, eye drops, gels, ointments, orally, intraocular injection,parenterally (e.g., intravenously or subcutaneous administration), byintramuscular injection, by intraperitoneal injection, subconjuctivalinjection, peri-/retrobulbar injection, transdermally, extracorporeally,by intracavity administration, transdermally, or topically or the like,including topical intranasal administration or administration byinhalant, and the like spray, emulsion, suspension, via any drugcarriers as sponges, contact lenses, polymers, microspheres, implants,pellets, and genetically engineered cells. The topical administrationcan be ophthalmically, vaginally, rectally, or intranasally. As usedherein, “topical intranasal administration” means delivery of thecompositions into the nose and nasal passages through one or both of thenares and can comprise delivery by a spraying mechanism or dropletmechanism, or through aerosolization of the nucleic acid or vector.Administration of the compositions by inhalant can be through the noseor mouth via delivery by a spraying or droplet mechanism. Delivery canalso be directly to any area of the respiratory system (e.g., lungs) viaintubation. The exact amount of the compositions required will vary fromsubject to subject, depending on the species, age, weight and generalcondition of the subject, the severity of the disorder being treated,the particular nucleic acid or vector used, its mode of administrationand the like. An appropriate amount for a particular composition and aparticular subject can be determined by one of ordinary skill in the artusing only routine experimentation given the teachings herein.

Parenteral administration of the composition, if used, is generallycharacterized by injection. Injectables can be prepared in conventionalforms, either as liquid solutions or suspensions, solid forms suitablefor solution of suspension in liquid prior to injection, or asemulsions. Parenteral administration includes use of a slow release, atime release or a sustained release system such that a constant dosageis maintained.

The active agent(s) can also be administered, as described herein, forimmediate delivery or extended release. The active agent(s) can also beadministered as dilutions, suspensions, emulsions, polymers,microspheres, gels, crèmes, and/or pellets. The active agents can alsobe administered in the form of drug carriers, sponges, polymersencapsulated cells, engineered cells, implants, and the like.

In one particularly preferred embodiment, the peptide(s) areadministered orally (e.g. as a syrup, capsule, or tablet).

The methods involve the administration of a single active agent of thisinvention or the administration of two or more different active agents.The active agents can be provided as monomers (e.g., in separate orcombined formulations), or in dimeric, oligomeric or polymeric forms. Incertain embodiments, the multimeric forms may comprise associatedmonomers (e.g., ionically or hydrophobically linked) while certain othermultimeric forms comprise covalently linked monomers (directly linked orthrough a linker).

While the invention is described with respect to use in humans, it isalso suitable for animal, e.g. veterinary use. Thus certain preferredorganisms include, but are not limited to humans, non-human primates,canines, equines, felines, porcines, ungulates, largomorphs, and thelike.

The methods of this invention are not limited to humans or non-humananimals showing one or more symptom(s) of the pathologies describedherein, but are also useful in a prophylactic context. Thus, the activeagents of this invention can be administered to organisms to prevent theonset/development of one or more symptoms of the pathologies describedherein (e.g., atherosclerosis, stroke, macular degeneration, etc.).Particularly preferred subjects in this context are subjects showing oneor more risk factors for the pathology. Thus, for example, in the caseof atherosclerosis risk factors include family history, hypertension,obesity, high alcohol consumption, smoking, high blood cholesterol, highblood triglycerides, elevated blood LDL, VLDL, IDL, or low HDL,diabetes, or a family history of diabetes, high blood lipids, heartattack, angina or stroke, etc.

In the case of eye disease, factors can include, but are not limited toage, family history, genetic predisposition, hypertension, obesity,cardiovascular health, fat intake, plasma lipids, oxidative stress,race, and exposure to sunlight.

P) Dosages

Effective dosages and schedules for administering the compositions maybe determined empirically, and making such determinations is within theskill in the art. The dosage ranges for the administration of thecompositions are those large enough to produce the desired effect inwhich the symptoms of the disorder are affected. The dosage should notbe so large as to cause adverse side effects, such as unwantedcross-reactions, anaphylactic reactions, and the like. Generally, thedosage will vary with the age, condition, sex and extent of the diseasein the patient, route of administration, or whether other drugs areincluded in the regimen, and can be determined by one of skill in theart. The dosage can be adjusted by the individual physician in the eventof any counter-indications. Dosage can vary, and can be administered inone or more dose administrations daily, for one or several days.Guidance can be found in the literature for appropriate dosages forgiven classes of pharmaceutical products. For example, disclosed hereinare methods comprising administering one or more of the disclosedsynthetic apolipoprotein E-mimicking peptides to a subject, wherebyplasma LDL, plasma VLDL, or both, are affected, wherein said syntheticapolipoprotein E-mimicking peptide is administered in an amount of about0.01 mg/kg to about 5 mg/kg.

Dosages also suitable for administration of the active agents disclosedherein include, but are not limited to dosages of 10 μg/ml to 400 μg/ml.For example, For example, disclosed herein are methods comprisingadministering one or more of the disclosed synthetic apolipoproteinE-mimicking peptides to a subject, whereby plasma LDL, plasma VLDL, orboth, are affected, wherein said synthetic apolipoprotein E-mimickingpeptide is administered in an amount of about 200 ug/ml to 800 ug/ml.

II. Active Agents

A wide variety of active agents are suitable for the treatment of one ormore of the indications discussed above. These agents include, but arenot limited to class A amphipathic helical peptides, class A amphipathichelical peptide mimetics of apoA-I having aromatic or aliphatic residuesin the non-polar face, small peptides including pentapeptides,tetrapeptides, tripeptides, dipeptides and pairs of amino acids, Apo-J(G* peptides), and peptide mimetics, e.g., as described below.

A) Class A Amphipathic Helical Peptides.

In certain embodiments, the activate agents for use in the method ofthis invention include class A amphipathic helical peptides, e.g. asdescribed in U.S. Pat. No. 6,664,230, and PCT Publications WO 02/15923and WO 2004/034977. It was discovered that peptides comprising a class Aamphipathic helix (“class A peptides”), in addition to being capable ofmitigating one or more symptoms of atherosclerosis are also useful inthe treatment of one or more of the other indications described herein.

Class A peptides are characterized by formation of an α-helix thatproduces a segregation of polar and non-polar residues thereby forming apolar and a nonpolar face with the positively charged residues residingat the polar-nonpolar interface and the negatively charged residuesresiding at the center of the polar face (see, e.g., Anantharamaiah(1986) Meth. Enzymol, 128: 626-668). It is noted that the fourth exon ofapo A-I, when folded into 3.667 residues/turn produces a class Aamphipathic helical structure.

One class A peptide, designated 18A (see, e.g., Anantharamaiah (1986)Meth. Enzymol, 128: 626-668) was modified as described herein to producepeptides orally administrable and highly effective at inhibiting orpreventing one or more symptoms of atherosclerosis and/or otherindications described herein. Without being bound by a particulartheory, it is believed that the peptides of this invention may act invivo by picking up seeding molecule(s) that mitigate oxidation of LDL.

Another theory could be that with macular degeneration, where thepresence of lipids in the Bruch's membrane causes the transfer of bloodfrom the eye vessels through the Bruch's membrane to the retinal pigmentcells and then to the photoreceptors to decrease. The decrease in bloodflow leads to a decrease in oxygen getting to the photoreceptors. Thebody then responds by creating more vasculature that invades the Bruch'smembrane and into the retinal pigment epithelial cells to compensate forthe decrease in oxygen supply to the photoreceptors and retinal pigmentepithelial cells. By providing one or more of the active agentsdescribed herein, the lipid accumulation could be removed and/orprevented, thereby relieving the need for increased vasculature. Inaddition, by providing one or more of the active agents described hereinin combination with an anti-angiogenic factor, not only could the lipidaccumulation be removed and/or prevented, the revascularization could beprevented as well, thereby relieving the need for increased vasculatureand preventing detrimental vascular growth.

We determined that increasing the number of Phe residues on thehydrophobic face of 18A would theoretically increase lipid affinity asdetermined by the computation described by Palgunachari et al. (1996)Arteriosclerosis, Thrombosis, & Vascular Biol. 16: 328-338.Theoretically, a systematic substitution of residues in the nonpolarface of 18A with Phe could yield six peptides. Peptides with anadditional 2, 3 and 4 Phe would have theoretical lipid affinity (X)values of 13, 14 and 15 units, respectively. However, the X valuesjumped four units if the additional Phe were increased from 4 to 5 (to19× units). Increasing to 6 or 7 Phe would produce a less dramaticincrease (to 20 and 21× units, respectively).

A number of these class A peptides were made including, the peptidedesignated 4F, D4F, 5F, and D5F, and the like. Various class A peptidesinhibited lesion development in atherosclerosis-susceptible mice. Inaddition, the peptides show varying, but significant degrees of efficacyin mitigating one or more symptoms of the various pathologies describedherein. A number of such peptides are illustrated in Table 2.

TABLE 2 Peptide Name Amino Acid Sequence SEQ ID NO. 18FD-W-L-K-A-F-Y-D-K-V-A-E-K-L-K-E-A-F 1 2FAc-D-W-L-K-A-F-Y-D-K-V-A-E-K-L-K-E-A-F-NH₂ 2 3FAc-D-W-F-K-A-F-Y-D-K-V-A-E-K-L-K-E-A-F-NH₂ 3 3F14Ac-D-W-L-K-A-F-Y-D-K-V-A-E-K-F-K-E-A-F-NH₂ 4 4FAc-D-W-F-K-A-F-Y-D-K-V-A-E-K-F-K-E-A-F-NH₂ 5 5FAc-D-W-L-K-A-F-Y-D-K-V-F-E-K-F-K-E-F-F-NH₂ 6 6FAc-D-W-L-K-A-F-Y-D-K-F-F-E-K-F-K-E-F-F-NH₂ 7 7FAc-D-W-F-K-A-F-Y-D-K-F-F-E-K-F-K-E-F-F-NH₂ 8Ac-D-W-L-K-A-F-Y-D-K-V-A-E-K-L-K-E-F-F-NH₂ 9Ac-D-W-L-K-A-F-Y-D-K-V-F-E-K-F-K-E-A-F-NH₂  10Ac-D-W-L-K-A-F-Y-D-K-V-F-E-K-L-K-E-F-F-NH₂ 11Ac-D-W-L-K-A-F-Y-D-K-V-A-E-K-F-K-E-F-F-NH₂ 12Ac-D-W-L-K-A-F-Y-D-K-V-F-E-K-F-K-E-F-F-NH₂ 13Ac-E-W-L-K-L-F-Y-E-K-V-L-E-K-F-K-E-A-F-NH₂ 14Ac-E-W-L-K-A-F-Y-D-K-V-A-E-K-F-K-E-A-F-NH₂  15Ac-E-W-L-K-A-F-Y-D-K-V-A-E-K-L-K-E-F-F-NH₂ 16Ac-E-W-L-K-A-F-Y-D-K-V-F-E-K-F-K-E-A-F-NH₂ 17Ac-E-W-L-K-A-F-Y-D-K-V-F-E-K-L-K-E-F-F-NH₂ 18Ac-E-W-L-K-A-F-Y-D-K-V-A-E-K-F-K-E-F-F-NH₂ 19Ac-E-W-L-K-A-F-Y-D-K-V-F-E-K-F-K-E-F-F-NH₂ 20Ac-A-F-Y-D-K-V-A-E-K-L-K-E-A-F-NH₂ 21 Ac-A-F-Y-D-K-V-A-E-K-F-K-E-A-F-NH₂22 Ac-A-F-Y-D-K-V-A-E-K-F-K-E-A-F-NH₂ 23Ac-A-F-Y-D-K-F-F-E-K-F-K-E-F-F-NH₂ 24 Ac-A-F-Y-D-K-F-F-E-K-F-K-E-F-F-NH₂25 Ac-A-F-Y-D-K-V-A-E-K-F-K-E-A-F-NH₂ 26Ac-A-F-Y-D-K-V-A-E-K-L-K-E-F-F-NH₂ 27 Ac-A-F-Y-D-K-V-F-E-K-F-K-E-A-F-NH₂28 Ac-A-F-Y-D-K-V-F-E-K-L-K-E-F-F-NH₂ 29Ac-A-F-Y-D-K-V-A-E-K-F-K-E-F-F-NH₂ 30 Ac-K-A-F-Y-D-K-V-F-E-K-F-K-E-F-NH₂31 Ac-L-F-Y-E-K-V-L-E-K-F-K-E-A-F-NH₂ 32Ac-A-F-Y-D-K-V-A-E-K-F-K-E-A-F-NH₂ 33 Ac-A-F-Y-D-K-V-A-E-K-L-K-E-F-F-NH₂34 Ac-A-F-Y-D-K-V-F-E-K-F-K-E-A-F-NH₂ 35Ac-A-F-Y-D-K-V-F-E-K-L-K-E-F-F-NH₂ 36 Ac-A-F-Y-D-K-V-A-E-K-F-K-E-F-F-NH₂37 Ac-A-F-Y-D-K-V-F-E-K-F-K-E-F-F NH₂ 38Ac-D-W-L-K-A-L-Y-D-K-V-A-E-K-L-K-E-A-L-NH₂ 39Ac-D-W-F-K-A-F-Y-E-K-V-A-E-K-L-K-E-F-F-NH₂ 40Ac-D-W-F-K-A-F-Y-E-K-F-F-E-K-F-K-E-F-F-NH₂ 41Ac-E-W-L-K-A-L-Y-E-K-V-A-E-K-L-K-E-A-L-NH₂ 42Ac-E-W-L-K-A-F-Y-E-K-V-A-E-K-L-K-E-A-F-NH₂ 43Ac-E-W-F-K-A-F-Y-E-K-V-A-E-K-L-K-E-F-F-NH₂ 44Ac-E-W-L-K-A-F-Y-E-K-V-F-E-K-F-K-E-F-F-NH₂ 45Ac-E-W-L-K-A-F-Y-E-K-F-F-E-K-F-K-E-F-F-NH₂ 46Ac-E-W-F-K-A-F-Y-E-K-F-F-E-K-F-K-E-F-F-NH₂ 47Ac-D-F-L-K-A-W-Y-D-K-V-A-E-K-L-K-E-A-W-NH₂ 48Ac-E-F-L-K-A-W-Y-E-K-V-A-E-K-L-K-E-A-W-NH₂ 49Ac-D-F-W-K-A-W-Y-D-K-V-A-E-K-L-K-E-W-W-NH₂ 50Ac-E-F-W-K-A-W-Y-E-K-V-A-E-K-L-K-E-W-W-NH₂ 51Ac-D-K-L-K-A-F-Y-D-K-V-F-E-W-A-K-E-A-F-NH₂ 52Ac-D-K-W-K-A-V-Y-D-K-F-A-E-A-F-K-E-F-L-NH₂ 53Ac-E-K-L-K-A-F-Y-E-K-V-F-E-W-A-K-E-A-F-NH₂ 54Ac-E-K-W-K-A-V-Y-E-K-F-A-E-A-F-K-E-F-L-NH₂ 55Ac-D-W-L-K-A-F-V-D-K-F-A-E-K-F-K-E-A-Y-NH₂ 56Ac-E-K-W-K-A-V-Y-E-K-F-A-E-A-F-K-E-F-L-NH₂ 57Ac-D-W-L-K-A-F-V-Y-D-K-V-F-K-L-K-E-F-F-NH₂ 58Ac-E-W-L-K-A-F-V-Y-E-K-V-F-K-L-K-E-F-F-NH₂ 59Ac-D-W-L-R-A-F-Y-D-K-V-A-E-K-L-K-E-A-F-NH₂ 60Ac-E-W-L-R-A-F-Y-E-K-V-A-E-K-L-K-E-A-F-NH₂ 61Ac-D-W-L-K-A-F-Y-D-R-V-A-E-K-L-K-E-A-F-NH₂ 62Ac-E-W-L-K-A-F-Y-E-R-V-A-E-K-L-K-E-A-F-NH₂ 63Ac-D-W-L-K-A-F-Y-D-K-V-A-E-R-L-K-E-A-F-NH₂ 64Ac-E-W-L-K-A-F-Y-E-K-V-A-E-R-L-K-E-A-F-NH₂ 65Ac-D-W-L-K-A-F-Y-D-K-V-A-E-K-L-R-E-A-F-NH₂ 66Ac-E-W-L-K-A-F-Y-E-K-V-A-E-K-L-R-E-A-F-NH₂ 67Ac-D-W-L-K-A-F-Y-D-R-V-A-E-R-L-K-E-A-F-NH₂ 68Ac-E-W-L-K-A-F-Y-E-R-V-A-E-R-L-K-E-A-F-NH₂ 69Ac-D-W-L-R-A-F-Y-D-K-V-A-E-K-L-R-E-A-F-NH₂ 70Ac-E-W-L-R-A-F-Y-E-K-V-A-E-K-L-R-E-A-F-NH₂ 71Ac-D-W-L-R-A-F-Y-D-R-V-A-E-K-L-K-E-A-F-NH₂ 72Ac-E-W-L-R-A-F-Y-E-R-V-A-E-K-L-K-E-A-F-NH₂ 73Ac-D-W-L-K-A-F-Y-D-K-V-A-E-R-L-R-E-A-F-NH₂ 74Ac-E-W-L-K-A-F-Y-E-K-V-A-E-R-L-R-E-A-F-NH₂ 75Ac-D-W-L-R-A-F-Y-D-K-V-A-E-R-L-K-E-A-F-NH₂ 76Ac-E-W-L-R-A-F-Y-E-K-V-A-E-R-L-K-E-A-F-NH₂ 77D-W-L-K-A-F-Y-D-K-V-A-E-K-L-K-E-A-F-P-D-W 78L-K-A-F-Y-D-K-V-A-E-K-L-K-E-A-FD-W-L-K-A-F-Y-D-K-V-A-E-K-L-K-E-F-F-P-D-W 79L-K-A-F-Y-D-K-V-A-E-K-L-K-E-F-FD-W-F-K-A-F-Y-D-K-V-A-E-K-L-K-E-A-F-P-D-W 80F-K-A-F-Y-D-K-V-A-E-K-L-K-E-A-FD-K-L-K-A-F-Y-D-K-V-F-E-W-A-K-E-A-F-P-D-K 81L-K-A-F-Y-D-K-V-F-E-W-L-K-E-A-FD-K-W-K-A-V-Y-D-K-F-A-E-A-F-K-E-F-L-P-D-K 82W-K-A-V-Y-D-K-F-A-E-A-F-K-E-F-LD-W-F-K-A-F-Y-D-K-V-A-E-K-F-K-E-A-F-P-D-W 83F-K-A-F-Y-D-K-V-A-E-K-F-K-E-A-FD-W-L-K-A-F-V-Y-D-K-V-F-K-L-K-E-F-F-P-D-W 84L-K-A-F-V-Y-D-K-V-F-K-L-K-E-F-FD-W-L-K-A-F-Y-D-K-F-A-E-K-F-K-E-F-F-P-D-W 85L-K-A-F-Y-D-K-F-A-E-K-F-K-E-F-FAc-E-W-F-K-A-F-Y-E-K-V-A-E-K-F-K-E-A-F-NH₂ 86Ac-D-W-F-K-A-F-Y-D-K-V-A-E-K-F-NH₂ 87 Ac-F-K-A-F-Y-D-K-V-A-E-K-F-K-E-NH₂88 Ac-F-K-A-F-Y-E-K-V-A-E-K-F-K-E-NH₂ 89NMA-F-K-A-F-Y-D-K-V-A-E-K-F-K-E-NH₂ 90NMA-F-K-A-F-Y-E-K-V-A-E-K-F-K-E-NH₂ 91NMA-D-W-F-K-A-F-Y-D-K-V-A-E-K-F-K-E-A-F-NH₂ 92NMA-E-W-F-K-A-F-Y-E-K-V-A-E-K-F-K-E-A-F-NH₂ 93NMA-A-F-Y-D-K-V-A-E-K-F-K-E-A-F-NH₂ 94NMA-D-W-F-K-A-F-Y-D-K-V-A-E-K-F-NH₂ 95Ac-D-W-L-K-A-F-Y-D-K-V-F-E-K-F-K-E-F-F-NH₂ 96NMA-D-W-L-K-A-F-Y-D-K-V-F-E-K-F-K-E-F-F-NH₂ 1198Ac-E-W-L- K-A-F-Y-E-K-V-F-E-K-F-K-E-F-F-NH₂ 97NMA-E-W-L-K-A-F-Y-E-K-V-F-E-K-F-K-E-F-F-NH₂ 1199Ac-A-F-Y-D-K-V-F-E-K-F-K-E-F-F-NH₂ 98NMA-A-F-Y-D-K-V-F-E-K-F-K-E-F-F-NH₂ 1200Ac-A-F-Y-E-K-V-F-E-K-F-K-E-F-F-NH₂ 99NMA-A-F-Y-E-K-V-F-E-K-F-K-E-F-F-NH₂ 1201Ac-D-W-L-K-A-F-Y-D-K-V-F-E-K-F-NH₂ 100NMA-D-W-L-K-A-F-Y-D-K-V-F-E-K-F-NH₂ 1202Ac-E-W-L-K-A-F-Y-E-K-V-F-E-K-F-NH₂ 101NMA-E-W-L-K-A-F-Y-E-K-V-F-E-K-F-NH₂ 1203Ac-L-K-A-F-Y-D-K-V-F-E-K-F-K-E-NH₂ 102NMA-L-K-A-F-Y-D-K-V-F-E-K-F-K-E-NH₂ 1204Ac-L-K-A-F-Y-E-K-V-F-E-K-F-K-E-NH₂ 103NMA-L-K-A-F-Y-E-K-V-F-E-K-F-K-E-NH₂ 1205 *Linkers are underlined. NMA isN-Methyl Anthranilyl.

In certain preferred embodiments, the peptides include variations of 4F((SEQ ID NO:5 in Table 2), also known as L-4F, where all residues are Lform amino acids) or D-4F where one or more residues are D form aminoacids). In any of the peptides described herein, the C-terminus, and/orN-terminus, and/or internal residues can be blocked with one or moreblocking groups as described herein.

While various peptides of Table 2, are illustrated with an acetyl groupor an N-methylanthranilyl group protecting the amino terminus and anamide group protecting the carboxyl terminus, any of these protectinggroups may be eliminated and/or substituted with another protectinggroup as described herein. In particularly preferred embodiments, thepeptides comprise one or more D-form amino acids as described herein. Incertain embodiments, every amino acid (e.g., every enantiomeric aminoacid) of the peptides of Table 2 is a D-form amino acid.

It is also noted that Table 2 is not fully inclusive. Using theteachings provided herein, other suitable class A amphipathic helicalpeptides can routinely be produced (e.g., by conservative orsemi-conservative substitutions (e.g., D replaced by E), extensions,deletions, and the like). Thus, for example, one embodiment utilizestruncations of any one or more of peptides shown herein (e.g., peptidesidentified by SEQ ID Nos:2-20 and 39—in Table 2). Thus, for example, SEQID NO:21 illustrates a peptide comprising 14 amino acids from theC-terminus of 18A comprising one or more D amino acids, while SEQ IDNOS:22-38 illustrate other truncations.

Longer peptides are also suitable. Such longer peptides may entirelyform a class A amphipathic helix, or the class A amphipathic helix(helices) can form one or more domains of the peptide. In addition, thisinvention contemplates multimeric versions of the peptides (e.g.,concatamers). Thus, for example, the peptides illustrated herein can becoupled together (directly or through a linker (e.g., a carbon linker,or one or more amino acids) with one or more intervening amino acids).Illustrative polymeric peptides include 18A-Pro-18A and the peptides ofSEQ ID NOs:78-85, in certain embodiments comprising one or more D aminoacids, more preferably with every amino acid a D amino acid as describedherein and/or having one or both termini protected.

It will also be appreciated in addition to the peptide sequencesexpressly illustrated herein, this invention also contemplates retro andretro-inverso forms of each of these peptides. In retro forms, thedirection of the sequence is reversed. In inverse forms, the chiralityof the constituent amino acids is reversed (i.e., L form amino acidsbecome D form amino acids and D form amino acids become L form aminoacids). In the retro-inverso form, both the order and the chirality ofthe amino acids is reversed. Thus, for example, a retro form of the 4Fpeptide (DWFKAFYDKVAEKFKEAF, SEQ ID NO:5), where the amino terminus isat the aspartate (D) and the carboxyl terminus is at the phenylalanine(F), has the same sequence, but the amino terminus is at thephenylalanine and the carboxy terminus is at the aspartate (i.e.,FAEKFKEAVKDYFAKFWD, SEQ ID NO:104). Where the 4F peptide comprises all Lamino acids, the retro-inverso form will have the sequence shown above(SEQ ID NO:104) and comprise all D form amino acids. As illustrated inthe helical wheel diagrams of FIG. 15, 4F and retroinverso (Rev-4F) aremirror images of each other with identical segregation of the polar andnonpolar faces with the positively charged residues residing at thepolar-nonpolar interface and the negatively charged residues residing atthe center of the polar face. These mirror images of the same polymer ofamino acids are identical in terms of the segregation of the polar andnonpolar faces with the positively charged residues residing at thepolar-nonpolar interface and the negatively charged residues residing atthe center of the polar face. For a discussion of retro- andretro-inverso peptides see, e.g., Chorev and Goodman, (1995) TibTech,13: 439-445.

Where reference is made to a sequence and orientation is not expresslyindicated, the sequence can be viewed as representing the amino acidsequence in the amino to carboxyl orientation, the retro form (i.e., theamino acid sequence in the carboxyl to amino orientation), the retroform where L amino acids are replaced with D amino acids or D aminoacids are replaced with L amino acids, and the retro-inverso form whereboth the order is reversed and the amino acid chirality is reversed.

C) Class A Amphipathic Helical Peptide Mimetics of apoA-1 havingAromatic or Aliphatic Residues in the Non-Polar Face.

In certain embodiments, this invention also provides modified class Aamphipathic helix peptides. Certain preferred peptides incorporate oneor more aromatic residues at the center of the nonpolar face, e.g.,3F^(cπ), (as present in 4F), or with one or more aliphatic residues atthe center of the nonpolar face, e.g., 3F^(Iπ), see, e.g., Table 3.Without being bound to a particular theory, we believe the centralaromatic residues on the nonpolar face of the peptide 3F^(Cπ), due tothe presence of π electrons at the center of the nonpolar face, allowwater molecules to penetrate near the hydrophobic lipid alkyl chains ofthe peptide-lipid complex, which in turn would enable the entry ofreactive oxygen species (such as lipid hydroperoxides) shielding themfrom the cell surface. Similarly, we also believe the peptides withaliphatic residues at the center of the nonpolar face, e.g., 3E^(Iπ),will act similarly but not quite as effectively as 3F^(Cπ).

Preferred peptides will convert pro-inflammatory HDL toanti-inflammatory HDL or make anti-inflammatory HDL moreanti-inflammatory, and/or decrease LDL-induced monocyte chemotacticactivity generated by artery wall cells equal to or greater than D4F orother peptides shown in Table 2.

TABLE 3 Name Sequence SEQ ID NO (3^(cn)) Ac-DKWKAVYDKFAEAFKEFL-NH₂ 105(3^(1n)) Ac-DKLKAFYDKVFEWAKEAF-NH₂ 106

D) Other Class A and Some Class Y Amphipathic Helical Peptides.

In certain embodiments this invention also contemplates class aamphipathic helical peptides that have an amino acid compositionidentical to one or more of the class a amphipathic helical peptidesdescribed above. Thus, for example, in certain embodiments thisinvention contemplates peptides having an amino acid compositionidentical to 4F. Thus, in certain embodiments, this invention includesactive agents that comprise a peptide that consists of 18 amino acids,where the 18 amino acids consist of 3 alanines (A), 2 aspartates (D), 2glutamates (E), 4 phenylalanines (F), 4 lysines (K), 1 valine (V), 1tryptophan (W), and 1 tyrosine (Y); and where the peptide forms a classA amphipathic helix; and protects a phospholipid against oxidation by anoxidizing agent. In various embodiments, the peptides comprise least one“D” amino acid residue; and in certain embodiments, the peptidescomprise all “D: form amino acid residues. A variety of such peptidesare illustrated in Table 4. Reverse (retro-), inverse, retro-inverso-,and circularly permuted forms of these peptides are also contemplated.

TABLE 4 Illustrative 18 amino acid length class A amphipathic helicalpeptides with the amino acid composition 3 alanines (A),2 aspartates (D), 2 glutamates (E), 4 phenylalanines (F),4 lysines (K), 1 valine (V), 1 tryptophan (W), and 1 tyrosine (Y). NameSequence SEQ ID NO [Switch D-E]-4F analogs 107 [Switch D-E]-1-4F Ac- EWFKAFY E KVA D KFK D AF-NH₂ 108 [Switch D-E]-2-4F Ac- E WFKAFYDKVADKFK EAF-NH₂ 109 [Switch D-E]-3-4F Ac-DWFKAFY E KVA D KFKEAF-NH₂ 110[Switch D-E]-4-4F Ac-DWFKAFY E KVAEKFK D AF-NH₂ 111[W-2, F-3 positions reversed] 112 4F-2 Ac-D FW KAFYDKVAEKFKEAF-NH₂ 113[Switch D-E]-1-4F-2 Ac- E FWKAFY E KVA D KFK D AF-NH₂ 114[Switch D-E]-2-4F-2 Ac- E FWKAFYDKVADKFK E AF-NH₂ 115[Switch D-E]-3-4F-2 Ac-DFWKAFY E KVA D KFKEAF-NH₂ 116[Switch D-E]-4-4F-2 Ac-DFWKAFY E KVAEKFK D AF-NH₂ 117[F-6 and Y-7 positions switched] 118 4F-3 Ac-DWFKA YF DKVAEKFKEAF-NH₂119 [Switch D-E]-1-4F-5 Ac- E WFKAYF E KVA D KFK D AF-NH₂ 120[Switch D-E]-2-4F-5 Ac- E WFKAYFDKVADKFK E AF-NH₂ 121[Switch D-E]-3-4F-5 Ac-DWFKAYF E KVA D KFKEAF-NH₂ 122[Switch D-E]-4-4F-5 Ac-DWFKAYF E KVAEKFK D AF-NH₂ 123[Y-land 10V positions switched] 124 4F-4 Ac-DWFKAF V DK Y AEKFKEAF-NH₂125 [Switch D-E]-1-4F-4 Ac- E WFKAFV E KYA D KFK D AF-NH₂ 126[Switch D-E]-2-4F-4 Ac- E WFKAFVDKYADKFK E AF-NH₂ 127[Switch D-E]-3-4F-4 Ac-DWFKAFV E KYA D KFKEAF-NH₂ 128 [Switch D-E]-4-4FAc-DWFKAFV E KYAEKFK D AF-NH₂ 129 [V-10 and A-11 switched] 130 4-F-5Ac-DWFKAFYDK AV EKFKEAF-NH₂ 131 [Switch D-E]-1-4F-5 Ac- E WFKAFY E KAV DKFK D AF-NH₂ 132 [Switch D-E]-2-4F-5 Ac- E WFKAFYDKAVDKFK E AF-NH₂ 133[Switch D-E]-3-4F-5 Ac-DWFKAFY E KAV D KFKEAF-NH₂ 134[Switch D-E]-4-4F-5 Ac-DWFKAFY E KAVEKFK D AF-NH₂ 135[A-11 and F-14 switched] 136 4F-6 Ac-DWFKAFYDKV F EK A KEAF-NH₂ 137[Switch D-E]-1-4F-6 Ac- E WFKAFY E KVF D KAKDAF-NH₂ 138[Switch D-E]-2-4F-6 Ac- E WFKAFYDKVFDKAKEAF-NH₂ 139 [Switch D-E]-3-4F-6Ac-DWFKAFY E KVF D KAK E AF-NH₂ 140 [Switch D-E]-4-4F-6 Ac-DWFKAFY EKVFEKAK D AF-NH₂ 141 [F-14 and A-17 switched] 142 4F-7 Ac-DWFKAFYDKVAEKA KE F F-NH₂ 143 [Switch D-E]-1-4F-7A Ac- E WFKAFYEKVA D KAK D FF-NH₂144 [Switch D-E]-2-4F-7 Ac- E WFKAFYDKVADKAK E FF-NH₂ 145[Switch D-E]-3-4F-7 Ac-DWFKAFYEKVA D KAKEFF-NH₂ 146 [Switch D-E]-4-4F-7Ac-DWFKAFYEKVAEKAK D FF-NH₂ 147 [A-17 and F-18 switched] 148 4F-8Ac-DWFKAFYDKVAEKFKE FA -NH₂ 149 [Switch D-E]-1-4F-8 Ac- E WFKAFY E KVA DKFK D FA-NH₂ 150 [Switch D-E]-2-4F-8 Ac- E WFKAFYDKVADKFK E FA-NH₂ 151[Switch D-E]-3-4F-8 Ac-DWFKAFY E KVA D KFKEFA-NH₂ 152[Switch D-E]-4-4F-8 Ac-DWFKAFY E KVAEKFK D FA-NH₂ 153[W-2 and A-17 switched] 154 4F-9 Ac-D A FKAFYDKVAEKFKE W F-NH₂ 155[Switch D-E]-1-4F-9 Ac- E AFKAFY E KVA D KFK D WF-NH₂ 156[Switch D-E]-2-4F-9 Ac- E AFKAFYDKVADKFK E WF-NH₂ 157[Switch D-E]-3-4F-9 Ac-DAFKAFY E KVA D KFKEWF-NH₂ 158[Switch D-E]-4-4F-9 Ac-DAFKAFY E KVAEKFK D WF-NH₂ 159[W-2 and A-11 switched] 160 4F-10 Ac-D A FKAFYDKV W EKFKEAF-NH₂ 161[Switch D-E]-1-4F-10 Ac- E AFKAFY E KVW D KFK D AF-NH₂ 162[Switch D-E]-2-4F-10 Ac- E AFKAFYDKVWDKFK E AF-NH₂ 163[Switch D-E]-3-4F-10 Ac-DAFKAFY E KVW D KFKEAF-NH₂ 164[Switch D-E]-4-4F-10 Ac-DAFKAFY E KVWEKFK D AF-NH₂ 165[W-2 and Y-7 switched] 166 4F-11 Ac-D Y FKAF W DKVAEKFKEAF-NH 167[Switch D-E]-1-4F-11 Ac- E YFKAFW E KVA D KFK D AF-NH₂ 168[Switch D-E]-2-4F-11 Ac- E YFKAFWDKVADKFK E AF-NH₂ 169[Switch D-E]-3-4F-11 Ac-DYFKAFW E KVA D KFKEAF-NH₂ 170[Switch D-E]-4-4F-11 Ac-DYFKAFW E KVAEKFK D AF-NH₂ 171[F-3 and A-17 switched] 172 4F-12 Ac-DWAK A FYDKVAEKFKE F F-NH₂ 173[Switch D-E]-1-4F-12 Ac- E WAKAFY E KVA D KFK D FF-NH₂ 174[Switch D-E]-2-4F-12 Ac- E WAKAFYDKVADKFK E FF-NH₂ 175[Switch D-E]-3-4F-12 Ac-DWAKAFY E KVA D KFKEFF-NH₂ 176[Switch D-E]-4-4F-12 Ac-DWAKAFY E KVAEKFK D FF-NH₂ 177[F-6 and A-17 switched] 178 4F-13 Ac-DWFK A AYDKVAEKFKE F F-NH₂ 179[Switch D-E]-1-4F-13 Ac- E WFKAAY E KVA D KFK D FF-NH₂ 180[Switch D-E]-2-4F-13 Ac- E WFKAAYDKVADKFK E FF-NH₂ 181[Switch D-E]-3-4F-13 Ac-DWFKAAY E KVA D KFKEFF-NH₂ 182[Switch D-E]-4-4F-13 Ac-DWFKAAY E KVAEKFK D FF-NH₂ 183[Y-7 and A-17 switched 184 4F-14 Ac-DWFK A FADKVAEKFKE Y F-NH₂ 185[Switch D-E]-1-4F-14 Ac- E WFKAFA E KVA D KFK D YF-NH₂ 186[Switch D-E]-2-4F-14 Ac- E WFKAFADKVADKFK E YF-NH₂ 187[Switch D-E]-3-4F-14 Ac-DWFKAFA E KVA D KFKEYF-NH₂ 188 [Switch D-E]-4-4F Ac-DWFKAFAEKVAEKFK D YF-NH₂ 189 [V-10 and A-17 switched] 190 4F-15Ac-DWFKAFYDK A AEKFKE V F-NH₂ 191 [Switch D-E]-1-4F-15 Ac- E WFKAFY EKAA D KFK D VF-NH₂ 192 [Switch D-E]-2-4F-15 Ac- E WFKAFYDKAADKFK EVF-NH₂ 193 [Switch D-E]-3-4F-15 Ac-DWFKAFY E KAA D KFKEVF-NH₂ 194[Switch D-E]-4-4F-15 Ac-DWFKAFY E KAAEKFK D VF-NH₂ 195[F3 and Y-7 switched] 196 4F-16 Ac-DW Y KAF F DKVAEKFKEAF-NH₂ 197[Switch D-E]-1-4F-16 Ac- E WYKAFF E KVA D KFK D AF-NH₂ 198[Switch D-E]-2-4F-16 Ac- E WYKAFFDKVADKFK E AF-NH₂ 199[Switch D-E]-3-4F-16 Ac-DWYKAFF E KVA D KFKEAF-NH₂ 200[Switch D-E]-4-4F-16 Ac-DWYKAFF E KVAEKFK D AF-NH₂ 201[F-3 and V-10 switched] 202 4F-17 Ac-DW V KAFYDK F AEKFKEAF-NH₂ 203[Switch D-E]-1-4F-17 Ac- E WVKAFY E KFA D KFK D AF-NH₂ 204[Switch D-E]-2-4F-17 Ac- E WVKAFYDKFADKFK E AF-NH₂ 205[Switch D-E]-3-4F-17 Ac-DWVKAFY E KFA D KFKEAF-NH₂ 206[Switch D-E]-4-4F-17 Ac-DWVKAFY E KFAEKFK D AF-NH₂ 207[Y-7 and F-14 switched] 208 4F-18 Ac-DWFKA F FDKVAEK Y KEAF-NH₂ 209[Switch D-E]-1-4F-18 Ac- E WFKAFF E KVA D KYK D AF-NH₂ 210[Switch D-E]-2-4F-18 Ac- E WFKAFFDKVADKYK E AF-NH₂ 211[Switch D-E]-3-4F-18 Ac-DWFKAFF E KVA D KYKEAF-NH₂ 212[Switch D-E]-4F-18 Ac-DWFKAFF E KVA D KYKEAF-NH₂ 213[Y-7 and F-18 switched] 214 4F-19 Ac-DWFKAF F DKVAEKFKEA Y -NH₂ 215[Switch D-E]-1-4F-19 Ac- E WFKAFF E KVA D KFK D AY-NH₂ 216[Switch D-E]-2-4F-19 Ac- E WFKAFFDKVADKFKEAY-NH₂ 217[Switch D-E]-3-4F-19 Ac-DWFKAFF E KVA D KFKEAY-NH₂ 218[Switch D-E]-4-4F-19 Ac-DWFKAFF E KVAEKFK D AY-NH₂ 219[V-10 and F-18 switched] 220 4F-20 Ac-DWFKAFYDK F AEKFKEA V -NH₂ 221[Switch D-E]-1-4F-20 Ac- E WFKAFY E KFA D KFK D AV-NH₂ 222[Switch D-E]-2-4F-20 Ac- E WFKAFYDKFADKFK E AV-NH₂ 223[Switch D-E]-3-4F-20  Ac-DWFKAFY E KFA D KFKEAV-NH₂ 224[Switch D-E]-4-4F-20 Ac-DWFKAFY E KFAEKFK D AV-NH₂ 225[W-2 and K13 switched] 226 4F-21 Ac-D K FKAFYDKVAEKF W EAF-NH₂ 227[Switch D-E]-1-4F-21 Ac- E KFKAFY E KVA D KFW D AF-NH₂ 228[Switch D-E]-2-4F-21 Ac- E KFKAFYDKVADKFW E AF-NH₂ 229[Switch D-E]-3-4F-21 Ac-DKFKAFY E KVA D KFWEAF-NH₂ 230[Switch D-E]-4-4F-21 Ac-DKFKAFY E KVAEKFW D AF-NH₂ 231[W-3, F-13 and K-2 4F] 232 4F-22 Ac-D KW KAYYDKVAEKF F EAF-NH₂ 233[Switch D-E]-1-4F-22 Ac- E KWKAFY E KVA D KFF D AF-NH₂ 234[Switch D-E]-2-4F-22 Ac- E KWKAFYDKVADKFF E AF-NH₂ 235[Switch D-E]-3-4F-22 Ac-DKWKAFY E KVA D KFFEAF-NH₂ 236[Switch D-E]-4-4F-22 Ac-DKWKAFY E KVAEKFF D AF-NH₂ 237 [K-2, W10, V-13]238 4F-23 Ac-DKF K AFYDK W AE V FKEAF-NH₂ 239 [Switch D-E]-4F analogs240 [Switch D-E]-1-4F-23 Ac- E KFKAFY E KWA D VFK D AF-NH₂ 241[Switch D-E]-2-4F-23 Ac- E KFKAFYDKWADVFK E AF-NH₂ 242[Switch D-E]-3-4F-23 Ac-DKFKAFY E KWA D VFKEAF-NH₂ 243[Switch D-E]-4-4F-23 Ac-DKFKAFY E KWAEVFK D AF-NH₂ 244[K-2, F-13, W-14 4F] 245 4F-24 Ac-D K FKAFYDKVAE FW KEAF-NH₂ 246[Switch D-E]-4F analogs 247 [Switch D-E]-1-4F-24 Ac- E KFKAFY E KVA DFWK D AF-NH₂ 248 [Switch D-E]-2-4F-24 Ac- E KFKAFYDKVADFWK E AF-NH₂ 249[Switch D-E]-3-4F-24 Ac-DKFKAFY E KVA D FWKEAF-NH₂ 250[Switch D-E]-4-4F-24 Ac-DKFKAFY E KVAEFWK D AF-NH₂ 251Reverse 4F analogs 252 Rev-4F Ac-FAEKFKEAVKDYFAKFWD-NH₂ 253[Switch D-E]-1-Rev-4F Ac-FA D KFK D AVK E YFAKFW E -NH₂ 254[Switch D-E]-2-Rev-4F Ac-FA D KFKEAVKDYFAKFW E -NH₂ 255[Switch D-E]-3-Rev-4F Ac-FAEKFK D AVK E YFAKFWD-NH₂ 256[Switch D-E]-4-Rev-4F Ac-FAEKFK D AVKDYFAKFW E -NH₂ 257[A-2 and W-17 switched] 258 Rev-4F-1 Ac-F W EKFKEAVKDYFAKF A D-NH₂ 259[Switch D-E]-1-Rev-4F-1 Ac-FW D KFK D AVK E YFAKFA E -NH₂ 260[Switch D-E]-2-Rev-4F-1 Ac-FA D KFKEAVKDYFAKFW E -NH₂ 261[Switch D-E]-3-Rev-4F-1 Ac-FAEKFK D AVK E YFAKFWD-NH₂ 262[Switch D-E]-4-Rev-4F-1 Ac-FAEKFK D AVKDYFAKFW E -NH₂ 263[Switch A-2 and F-16] 264 Rev-4F-2 Ac-F F EKFKEAVKDYFAK A WD-NH₂ 265[Switch D-E]-1-Rev-4F-2 Ac-FF D KFK D AVK E YFAKAW E -NH₂ 266[Switch D-E]-2-Rev-4F-2 Ac-FF D KFKEAVKDYFAKAW E -NH₂ 267[Switch D-E]-3-Rev-4F-2 Ac-FFEKFK D AVK E YFAKAWD-NH₂ 268[Switch D-E]-4-Rev-4F-2 Ac-FFEKFK D AVKDYFAKAW E -NH₂ 269[switch F-5 and A-8] 270 Rev-4F-3 Ac-FAEK A KE F VKDYFAKFWD-NH₂ 271[Switch D-E]-1-Rev-4F-3 Ac-FA D KAK D FVK E YFAKFW E -NH₂ 272[Switch D-E]-2-Rev-4F-3 Ac-FA D KAKEFVKDYFAKFW E -NH₂ 273[Switch D-E]-3-Rev-4F-3 Ac-FAEKAK D FVK E YFAKFWD-NH₂ 274[Switch D-E]-4-Rev-4F-3 Ac-FAEKAK D FVKDYFAKFW E -NH₂ 275[Switch A-8 and V9] 276 Rev-4F-4 Ac-FAEKFKE VA KDYFAKFWD-NH₂ 277[Switch D-E]-1-Rev-4F-4 Ac-FA D KFK D VAK E YFAKFW E -NH₂ 278[Switch D-E]-2-Rev-4F-4 Ac-FA D KFKEVAKDYFAKFW E -NH₂ 279[Switch D-E]-3-Rev-4F-4 Ac-FAEKFK D VAK E YFAKFWD-NH₂ 280[Switch D-E]-4-Rev-4F-4 Ac-FAEKFK D VAKDYFAKFW E -NH₂ 281[Switch V-9 to Y-12] 282 Rev-4F-5 Ac-FAEKFKEA Y KD V FAKFWD-NH₂ 283[Switch D-E]-1-Rev-4F-5 Ac-FA D KFK D AYK E VFAKFW E -NH₂ 284[Switch D-E]-2-Rev-4F-5 Ac-FA D KFKEAYKDVFAKFW E -NH₂ 285[Switch D-E]-3-Rev-4F-5 Ac-FAEKFK D AYK E VFAKFWD-NH₂ 286[Switch D-E]-4-Rev-4F-5 Ac-FAEKFK D AYKDVFAKFW E -NH₂ 287[Switch Y-12 and F-13] 288 Rev-4F-6 Ac-FAEKFKEAVKD FY AKFWD-NH₂ 289[Switch D-E]-1-Rev-4F-6 Ac-FA D KFK D AVK E FYAKFW E -NH₂ 290[Switch D-E]-2-Rev-4F-6 Ac-FA D KFKEAVKDFYAKFW E -NH₂ 291[Switch D-E]-3-Rev-4F-6 Ac-FAEKFK D AVK E FYAKFWD-NH₂ 292[Switch D-E]-4-Rev-4F-6 Ac-FAEKFK D AVKDFYAKFW E -NH₂ 293[Switch K-6 and W-17] 294 Rev-4F-7 Ac-FAEKF W EAVKDYFAKF K D-NH₂ 295[Switch D-E]-1-Rev-4F-7 Ac-FA D KFW D AVK E YFAKFK E -NH₂ 296[Switch D-E]-2-Rev-4F-7 Ac-FA D KFWEAVKDYFAKFK E -NH₂ 297[Switch D-E]-3-Rev-4F-7 Ac-FAEKFW D AVK E YFAKFKD-NH₂ 298[Switch D-E]-4-Rev-4F-7 Ac-FAEKFW D AVKDYFAKFK E -NH₂ 299[Switch F-1 and A-2] 300 Rev-4F-8 Ac- AF EKFKEAVKDYFAKFWD-NH₂ 301[Switch D-E]-1-Rev-4F-8 Ac-AF D KFK D AVK E YFAKFW E -NH₂ 302[Switch D-E]-2-Rev-4F-8 Ac-AF D KFKEAVKDYFAKFW E -NH₂ 303[Switch D-E]-3-Rev-4F-8 Ac-AFEKFK D AVK E YFAKFWD-NH₂ 304[Switch D-E]-4-Rev-4F-8 Ac-AFEKFK D AVKDYFAKFW E -NH₂ 305[F-1 and V-9 are switched] 306 Rev-F-9 Ac- V AEKFKEA F KDYFAKFWD-NH₂ 307[Switch D-E]-1-Rev-4F-9 Ac-VA D KFK D AFK E YFAKFW E -NH₂ 308[Switch D-E]-2-Rev-4F-9 Ac-VA D KFKEAFKDYFAKFW E -NH₂ 309[Switch D-E]-3-Rev-4F-9 Ac-VAEKFK D AFK E YFAKFWD-NH₂ 310[Switch D-E]-4-Rev-4F-9 Ac-VAEKFK D AFKDYFAKFW E -NH₂ 311[F-1 and Y-12 are switched] 312 Rev-4F-10 Ac- Y AEKFKEAVKD F FAKFWD-NH₂313 [Switch D-E]-1-Rev-4F-10 Ac-YA D KFK D AVK E FFAKFW E -NH₂ 314[Switch D-E]-2-Rev-4F-10 Ac-YA D KFKEAVKDFFAKFW E -NH₂ 315[Switch D-E]-3-Rev-4F-10 Ac-YAEKFK D AVK E FFAKFWD-NH₂ 316[Switch D-E]-4-Rev-4F-10 Ac-YAEKFK D AVKDFFAKFW E -NH₂ 317[F-1 and A-8 are switched] 318 Rev-4F-11 Ac- A AEKFKE F VKDYFAKFWD-NH₂319 [Switch D-E]-1-Rev-4F-11 Ac-AA D KFK D FVK E YFAKFW E -NH₂ 320[Switch D-E]-2-Rev-4F-11 Ac-AA D KFKEFVKDYFAKFW E -NH₂ 321[Switch D-E]-3-Rev-4F-11 Ac-AAEKFK D FVK E YFAKFWD-NH₂ 322Switch D-E]-4-Rev-4F-11 Ac-AAEKFKDFVK D YFAKFW E -NH₂ 323[A-2 and F-5 are switched] 324 Rev-4F-12 Ac-F F EK A KEAVKDYFAKFWD-NH₂325 [Switch D-E]-1-Rev-4F-12 Ac-FF D KAK D AVK E YFAKFW E -NH₂ 326[Switch D-E]-2-Rev-4F-12 Ac-FF D KAKEAVKDYFAKFW E -NH₂ 327[Switch D-E]-3-Rev-4F-12 Ac-141-EKAK D AVK E YFAKFWD-NH₂ 328[Switch D-E]-4-Rev-4F-12 Ac-1-1-EKAK D AVKDYFAKFW E -NH₂ 329[A-2 and Y12 are switched 330 Rev-4F-13 Ac-F Y EKFKEAVKD A FAKFWD-NH₂331 [Switch D-E]-1-Rev-4F-13 Ac-FY D KFK D AVK E AFAKFW E -NH₂ 332[Switch D-E]-2-Rev-4F-13 Ac-FY D KFKEAVKDAFAKFW E -NH₂ 333[Switch D-E]-3-Rev-4F-13 Ac-FYEKFK D AVK E AFAKFWD-NH₂ 334[Switch D-E]-4-Rev-4F-13 Ac-FYEKFK D AVKDAFAKFW E -NH₂ 335[A-2 and V-9 are switched] 336 Rev-4F-14 Ac-F V EKFKEA A KDYFAKFWD-NH₂337 [Switch D-E]-1-Rev-4F-14 Ac-FV D KFK D AAK E YFAKFW E -NH₂ 338[Switch D-E]-2-Rev-4F-14 Ac-FV D KFKEAAKDYFAKFW E -NH₂ 339[Switch D-E]-3-Rev-4F-14 Ac-FVEKFK D AAK E YFAKFWD-NH₂ 340[Switch D-E]-4-Rev-4F-14 Ac-FVEKFK D AAKDYFAKFW E -NH₂ 341[F-5 and Y-12 are switched] 342 Rev-4F-15 Ac-FAEK Y KEAVKD F FAKFWD-NH₂343 [Switch D-E]-1-Rev-4F-15 Ac-FA D KYK D AVK E FFAKFW E -NH₂ 344[Switch D-E]-2-Rev-4F-15 Ac-FA D KYKEAVKDFFAKFW E -NH₂ 345[Switch D-E]-3-Rev-4F-15 Ac-FAEKYK D AVK E FFAKFWD-NH₂ 346[Switch D-E]-4-Rev-4F-15 Ac-FAEKYK D AVKDFFAKFW E -NH₂ 347[F-5 and V-9 are switched] 348 Rev-4F-16 Ac-FAEK V KEA F KDYFAKFWD-NH₂349 [Switch D-E]-1-Rev-4F-16 Ac-FA D KVK D AFK E YFAKFW E -NH₂ 350[Switch D-E]-2-Rev-4F-16 Ac-FA D KVKEAFKDYFAKFW E -NH₂ 351[Switch D-E]-3-Rev-4F-16 Ac-FAEKVK D AFK E YFAKFWD-NH₂ 352[Switch D-E]-4-Rev-4F-16 Ac-FAEKVK D AFKDYFAKFW E -NH₂ 353[A-8 and Y-12 switched] 354 Rev-4F-17 Ac-FAEKFKE Y VKD A FAKFWD-NH₂ 355[Switch D-E]-1-Rev-4F-17 Ac-FA D KFK D YVK E AFAKFW E -NH₂ 356[Switch D-E]-2-Rev-4F-17 Ac-FA D KFKEYVKDAFAKFW E -NH₂ 357[Switch D-E]-3-Rev-4F-17 Ac-FAEKFK D YVK E AFAKFWD-NH₂ 358[Switch D-E]-4-Rev-4F-17 Ac-FAEKFK D YVKDAFAKFW E -NH₂ 359[V-9 and F-13 are switched] 360 Rev-4F-18 Ac-FAEKFKEA F KDYVAKFWD-NH₂361 [Switch D-E]-1-Rev-4F-18 Ac-FA D KFK D AFK E YVAKFW E -NH₂ 362[Switch D-E]-2-Rev-4F-18 Ac-FA D KFKEAFKDYVAKFW E -NH₂ 363[Switch D-E]-3-Rev-4F-18 Ac-FAEKFK D AFK E YVAKFWD-NH₂ 364[Switch D-E]-4-Rev-4F-18 Ac-FAEKFK D AFKDYVAKFW E -NH₂ 365[V-9 and F-16 switched] 366 Rev-4F-19 Ac-FAEKFKEA F KDYEAK V WD-NH₂ 367[Switch D-E]-1-Rev-4F-19 Ac-FA D KFK D AFK E YFAKVW E -NH₂ 368[Switch D-E]-2-Rev-4F-19 Ac-FA D KFKEAFKDYFAKVW E -NH₂ 369[Switch D-E]-3-Rev-4F-19 Ac-FAEKFK D AFK E YFAKVWD-NH₂ 370Switch D-E]-4-Rev-4F-19 Ac-FAEKFK D AFKDYFAKVW E -NH₂ 371[Y-12 and F-16 are switched 372 Rev-4F-20 Ac-FAEKFKEAVKD F FAK Y WD-NH₂373 [Switch D-E]-1-Rev-4F-20 Ac-FA D KFK D AVK E FFAKYW E -NH₂ 374[Switch D-E]-2-Rev-4F-20 Ac-FA D KFKEAVKDFFAKYW E -NH₂ 375[Switch D-E]-3-Rev-4F-20 Ac-FAEKFK D AVK E FFAKYWD-NH₂ 376[Switch D-E]-4-Rev-4F-20 Ac-FAEKFK D AVKDFFAKYW E -NH₂ 377[W-1, F-6 and K-17 Rev 4F] 378 Rev-4F-21 Ac- W AEKF F EAVKDYFAKF K D-NH₂379 [Switch D-E]-1-Rev-4F-7 Ac-WA D KFF D AVK E YFAKFK E -NH₂ 380[Switch D-E]-2-Rev-4F-7 Ac-WA D KFFEAVKDYFAKFK E -NH₂ 381[Switch D-E]-3-Rev-4F-7 Ac-WAEKFF D AVK E YFAKFKD-NH₂ 382Switch D-E]-4-Rev-4F-7 Ac-WAEKFF D AVKDYFAKFK E -NH₂ 383[W-5, F-6 and K-17 Rev-4F] 384 Rev-4F-22 Ac-FAEK WF EAVKDYFAKF K D-NH₂385 [Switch D-E]-1-Rev-4F-22 Ac-FA D KWF D AVK E YFAKFK E -NH₂ 386[Switch D-E]-2-Rev-4F-22 Ac-FA D KWFEAVKDYFAKFK E -NH₂ 387[Switch D-E]-3-Rev-4F-22 Ac-FAEKWF D AVK E YFAKFKD-NH₂ 388[Switch D-E]-4-Rev-4F-22 Ac-FAEKWF D AVKDYFAKFK E -NH₂ 389[V-6, W-9, K-17 Rev-4F] 390 Rev-4F-23 Ac-FAEKF V EA W KDYFAKF K D-NH₂391 [Switch D-E]-1-Rev-4F-23 Ac-FA D KFV D AWK E YFAKFK E -NH₂ 392[Switch D-E]-2-Rev-4F-23 Ac-FA D KFVEAWKDYFAKFK E -NH₂ 393[Switch D-E]-3-Rev-4F-23 Ac-FAEKFV D AWK E YFAKFKD-NH₂ 394[Switch D-E]-4-Rev-4F-23 Ac-FAEKFV D AWKDYFAKFK E -NH₂ 395[Y-2, A-4, W-12, K-17 Rev-4F] 396 Rev-4F-24 Ac-F Y EKF A EAVKD W FAKF KD-NH₂ 397 [Switch D-E]-1-Rev-4F-24 Ac-FY D KFA D AVK E WFAKFKE-NH₂ 398[Switch D-E]-2-Rev-4F-24 Ac-FY D KFAEAVKDWFAKFK E -NH₂ 399[Switch D-E]-3-Rev-4F-24 Ac-FYEKFA D AVK E WFAKFKD-NH₂ 400[Switch D-E]-4-Rev-4F-24 Ac-FYEKFA D AVKDWFAKFK E -NH₂ 401

Based on the helical wheel diagrams shown in FIG. 15 it is possible toreadily identify biologically active and useful peptides. Thus, forexample, the following peptides have been accurately identified asactive: 3F1; 3F2; 4F the reverse (retro) forms thereof and theretro-inverso forms thereof. Thus, in certain embodiments, thisinvention contemplates active agents comprising a peptide that is 18amino acids in length and forms a class A amphipathic helix where thepeptide has the amino acid composition 2 aspartates, 2 glutamates, 4lysines, 1 tryptophan, 1 tyrosine, no more than one leucine, no morethan 1 valine, no less than 1 and no more than 3 alanines, and with 3 to6 amino acids from the group: phenylalanine, alpha-naphthalanine,beta-naphthalanine, histidine, and contains either 9 or 10 amino acidson the polar face in a helical wheel representation of the class Aamphipathic helix including 4 amino acids with positive charge atneutral pH with two of the positively charged residues residing at theinterface between the polar and non-polar faces and with two of the fourpositively charged residues on the polar face that are contiguous and onthe non-polar face two of the amino acid residues from the group:phenylalanine, alpha-naphthalanine, beta-naphthalanine, histidine arealso contiguous and if there are 4 or more amino acids from this groupon the non-polar face there are also at least 2 residues from this groupthat are not contiguous.

In certain embodiments, this invention also contemplates certain class Yas well as class A amphipathic helical peptides. Class Y amphipathichelical peptides are known to those of skill in the art (see, e.g.,Segrest et al. (1992) J. Lipid Res. 33: 141-166; Oram and Heinecke(2005) Physiol Rev. 85: 1343-1372, and the like). In various embodimentsthese peptides include, but are not limited to an 18 amino acid peptidethat forms a class A amphipathic helix or a class Y amphipathic helixdescribed by formula III (SEQ ID NO:402):

D X X K Y X X D K X Y D KX K D Y X III

where the D's are independently Asp or Glu; the Ks are independently Lysor Arg; the Xs are independently Leu, norLeu, Val, Ile, Trp, Phe, Tyr,β-Nal, or α-Nal and all X residues are on the non-polar face (e.g., whenviewed in a helical wheel diagram) except for one that can be on thepolar face between two K residues; the Y's are independently Ala, His,Ser, Gln, Asn, or Thr non-polar face (e.g., when viewed in a helicalwheel diagram) and the Y's are independently one Ala on the polar face,one His, one Ser, one Gln one Asn, or one Thr on the polar face (e.g.,when viewed in a helical wheel diagram), where no more than two K are becontiguous (e.g., when viewed in a helical wheel diagram); and where nomore than 3 D's are contiguous (e.g., when viewed in a helical wheeldiagram) and the fourth D is be separated from the other D's by a Y.Illustrative peptides of this kind which include peptides withhistidine, and/or alpha- and/or beta-napthalanine are shown in Table 5.Reverse (retro-), inverse, retro-inverso-, and circularly permuted formsof these peptides are also contemplated.

TABLE 5 SEQ ID Short Name Peptide Sequence NO. [A-5 > H]4FAc-DWFKHFYDKVAEKFKEAF-NH₂ 403 [A-5 > H, D-E switched] 4FAc-EWFKHFYEKVADKFKDAF-NH₂ 404 [A-5 > H, D-1 > E]4FAc-EWFKHFYDKVAEKFKEAF-NH₂ 405 [A-5 > H, D-8 > E]4-FAc-DWFKHFYEKVAEKFKEAF-NH₂ 406 [A-5 > H, E-12 > D] 4FAc-DWFKHFYDKVADKFKEAF-NH₂ 407 [A-5 > H, E-16 > D]4FAc-DWFKHFYDKVAEKFKDAF-NH₂ 408 [F-3 > H, A-5 > F]-4FAc-DWHKFFYDKVAEKFKEAF-NH₂ 409 [F-3 > H, A-5 > F, D-E switched]-4FAc-EWHKFFYEKVADKFKDAF-NH₂ 410 [F-3 > H, A-5 > F, D-1 > E]-4FAc-EWHKFFYDKVAEKFKEAF-NH₂ 411 [F-3 > H, A-5 > F, D-8 > E]-4FAc-DWHKFFYEKVAEKFKEAF-NH₂ 412 [F-3 > H, A-5 > F, E- 12 > D]-4FAc-DWHKFFYDKVADKFKEAF-NH₂ 413 [F-3 > H, A-5 > F, E-16 > D]-4FAc-DWHKFFYDKVAEKFKDAF-NH₂ 414 [A-5 > F, F-6 > H]4FAc-DWFKFHYDKVAEKFKEAF-NH₂ 415 [A-5 > F, F-6 > H, D-E switched]4FAc-EWFKFHYEKVADKFKDAF-NH₂ 416 [[A-5 > F, F-6 > H, D- 1 > E]4FAc-EWFKFHYDKVAEKFKEAF-NH₂ 417 [A-5 > F, F-6 > H, D-8 > E]4FAc-DWFKFHYEKVAEKFKEAF-NH₂ 418 [A-5 > F, F-6 > H, E-12 > D]4FAc-DWFKFHYDKVADKFKEAF-NH₂ 419 [A-5 > F, F-6 > H, E- 16 > D]4FAc-DWFKFHYDKVAEKFKDAF-NH₂ 420 [A-5 > V, V-10 > H]4FAc-DWFKVFYDKHAEKFKEAF-NH₂ 421 [A-5 > V, V-10 > H, D-E switched]4FAc-EWFKVFYEKHADKFKDAF-NH₂ 422 [A-5 > V, V-10 > H, D-1 > E]4FAc-EWFKVFYDKHAEKFKEAF-NH₂ 423 [A-5 > V, V-10 > H, D-8 > E]4FAc-DWFKVFYEKHAEKFKEAF-NH₂ 424 [A-5 > V, V-10 > H, E-12 > D]4FAc-DWFKVFYDKHADKFKEAF-NH₂ 425 [A-5 > V, V-10 > H, E16 > D]4FAc-DWFKVFYDKHAEKFKDAF-NH₂ 426 [[A-17 > H]4F Ac-DWFKAFYDKVAEKFKEHF-NH₂427 [A-17 > H, D-E switched]4F Ac-EWFKAFYEKVADKFKDHF-NH₂ 428 [[A-17 >H, D-1 > E]4F Ac-EWFKAFYDKVAEKFKEHF-NH₂ 429 [[A-17 > H, D-8 > E]4FAc-DWFKAFYEKVAEKFKEHF-NH₂ 430 [[A-17 > H, E-12 > D]4FAc-DWFKAFYDKVADKFKEHF-NH₂ 431 [[A-17 > H, E 1 6 > D]4FAc-DWFKAFYDKVAEKFKDHF-NH₂ 432 [A-17 > F, F-18 > H]4FAc-DWFKAFYDKVAEKFKEFH-NH₂ 433 [A-17 > F, F-18 > H, D-E switched]4FAc-EWFKAFYEKVADKFKDFH-NH₂ 434 [A-17 > F, F-18 > H, D-1 > E]-4FAc-EWFKAFYDKVAEKFKEFH-NH₂ 435 [A-17 > F, F-18 > H]4FAc-DWFKAFYDKVAEKFKEFH-NH₂ 436 [A-17 > F, F-18 > H, D-8 > E]-4FAc-DWFKAFYEKVAEKFKEFH-NH₂ 437 [A-17 > F, F-18 > H, E-12 > D]4FAc-DWFKAFYDKVAEKFKEFH-NH₂ 438 [A-17 > F, F-18 > H], E-16 > D]-4FAc-DWFKAFYDKVAEKFKDFH-NH₂ 439 Rev-4F Ac-FAEKFKEAVKDYFAKFWD-NH₂ 440[A-2 > H]Rev4F Ac-FHEKFKEAVKDYFAKFWD-NH₂ 441 Rev-[A-2 > H, D > E]-4FAc-FHEKFKEAVKEYFAKFWE-NH₂ 442 Rev-[A-2 > H, E]22D]4FAc-FHDKFKDAVKDYFAKFWD-NH₂ 443 [A-2 > H, D-E switched]Rev-4FAc-FHDKFKDAVKEYFAKFWE-NH₂ 444 [A-2 > H, E-3 > D]Rev-4FAc-FHDKFKEAVKDYFAKFWD-NH₂ 445 [A-2 > H, E-7 > D]Rev-4FAc-FHEKFKDAVKDYFAKFWD-NH₂ 446 [A-2 > 2H, D-11 > E]Rev-4FAc-FHEKFKEAVKEYFAKFWD-NH₂ 447 [A-2 > H, D-18 > E]Rev-4FAc-FHEKFKEAVKDYFAKFWE-NH₂ 448 [F-1 > H, A-2 > F+ Rev-4FAc-HFEKFKEAVKDYFAKFWD-NH₂ 449 [F-1 > H, A-2 > F,D-E switched]Rev-4FAc-HFDKFKDAVKEYFAKFWE-NH₂ 450 [F-1 > H, A-2 > F, D > E]Rev-4FAc-HFEKFKEAVKEYFAKFWE-NH₂ 451 [F-1 > H, A-2 > F, E-3 > D]Rev-4FAc-HFDKFKEAVKDYFAKFWD-NH₂ 452 [F-1 > H, A-2 > F, E-7 > D]Rev-4FAc-HFEKFKDAVKDYFAKFWD-NH₂ 453 [F-1 > H, A-2 > F, D-11 > E]Rev-4FAc-HFEKFKEAVKEYFAKFWD-NH₂ 454 [F-1 > H, A-2 > F, D-18 > E]Rev-4FAc-HFEKFKEAVKDYFAKFWE-NH₂ 455 [A-2 > F, F-5 > H] Rev D-4FAc-FFEKHKEAVKDYFAKFWD-NH₂ 456 [A-2 > F, F-5 > H, D-E switched]Rev D-4FAc-FFDKHKDAVKEYFAKFWE-NH₂ 457 [A-2 > F, F-5 > H, D > E]Rev D-4FAc-FFEKHKEAVKEYFAKFWE-NH₂ 458 [A-2 > F, F-5 > H, E]22D]Rev D-4FAc-FFDKHKDAVKDYFAKFWD-NH₂ 459 A-2 > F, F-5 > H, E-3 > D]RevAc-FFDKHKEAVKDYFAKFWD-NH₂ 460 D-4F [A-2 > F, F-5 > H, D-11 > E]Rev D-4FAc-FFEKHKEAVKEYFAKFWD-NH₂ 461 [A-2 > F, F-5 > H, D-18 > E]Rev D-4FAc-FFEKHKEAVKDYFAKFWE-NH₂ 462 [A-2 > V, V-9 > H]Rev D-4FAc-FVEKFKEAHKDYFAKFWD-NH₂ 463 [A-2 > V, V-9 > H, D- E switched]Rev D-4FAc-FVDKFKDAHKEYFAKFWE-NH₂ 464 [A-2 > V, V-9 > H, D > E]Rev D-4FAc-FVEKFKEAHKEYFAKFWE-NH₂ 465 [A-2 > V, V-9 > H, E]22D]Rev D-4FAc-FVDKFKDAHKDYFAKFWD-NH₂ 466 [A-2 > V, V-9 > H, E-3 > D]Rev D-4FAc-FVDKFKEAHKDYFAKFWD-NH₂ 467 [A-2 > V, V-9 > H, E-7 > D]Rev D-4FAc-FVEKFKDAHKDYFAKFWD-NH₂ 468 [A-2 > V, V-9 > H, D-11 > E]Rev D-4FAc-FVEKFKEAHKEYFAKFWD-NH₂ 469 [A-2 > V, V-9 > H, D-18 > E]Rev D-4FAc-FVEKFKEAHKDYFAKFWE-NH₂ 470 [A-8 > H]Rev-4F Ac-FAEKFKEHVKDYFAKFWD-NH₂471 [A-8 > H,D-E switched]Rev-4F Ac-FADKFKDHVKEYFAKFWE-NH₂ 472 [A-8 >H, D > E]Rev-4F Ac-FAEKFKEHVKEYFAKFWE-NH₂ 473 [A-8 > H, E]22D]Rev-4FAc-FADKFKDHVKDYFAKFWD-NH₂ 474 [A-8 > H, E-3 > D]Rev-4FAc-FADKFKEHVKDYFAKFWD-NH₂ 475 [A-8 > H, E-7 > D]Rev-4FAc-FAEKFKDHVKDYFAKFWD-NH₂ 476 [A-8 > H, D-11 > E]Rev-4FAc-FAEKFKEHVKEYFAKFWD-NH₂ 477 [A-8 > H, D-18 > E] Rev-4FAc-FAEKFKEHVKDYFAKFWE-NH₂ 478 [A-8 > F, F-13 > H]Rev-4FAc-FAEKFKEFVKDYHAKFWD-NH₂ 479 [A-8 > F, F-13 > H, D-E switched]Rev-4FAc-FADKFKDFVKEYHAKFWE-NH₂ 480 [A-8 > F, F-13 > H, E-3 > D]Rev-4FAc-FADKFKEFVKDYHAKFWD-NH₂ 481 [A-8 > F, F-13 > H, E-7 > D]Rev-4FAc-FAEKFKDFVKDYHAKFWD-NH₂ 482 [A-8 > F, F-13 > H, E]22D]Rev-4FAc-FADKFKDFVKDYHAKFWD-NH₂ 483 [A-8 > F, F-13 > H, D > E]Rev-4FAc-FAEKFKEFVKEYHAKFWE-NH₂ 484 [A-8 > F, F-13 > H, D-11 > E]Rev-4FAc-FAEKFKEFVKEYHAKFWD-NH₂ 485 [A-8 > F, F-13 > H, D-18 > E]Rev-4FAc-FAEKFKEFVKDYHAKFWE-NH₂ 486 [A-8 > F, F16 > H]Rev-4FAc-FAEKFKEFVKDYFAKHWD-NH₂ 487 [A-8 > F, F16 > H, D-E switched]Rev-4FAc-FADKFKDFVKEYFAKHWE-NH₂ 488 [A-8 > F, F16 > H, D > E]Rev-4FAc-FAEKFKEFVKEYFAKHWE-NH₂ 489 [A-8 > F, F16 > H, E]22D]Rev-4FAc-FADKFKDFVKDYFAKHWD-NH₂ 490 [A-8 > F, F16 > H, E- 3 > D]Rev-4FAc-FADKFKEFVKDYFAKHWD-NH₂ 491 [A-8 > F, F16 > H, E-7 > D]Rev-4FAc-FAEKFKDFVKDYFAKHWD-NH₂ 492 [A-8 > F, F16 > H, D- 11 > E]Rev-4FAc-FAEKFKEFVKEYFAKHWD-NH₂ 493 [A-8 > F, F16 > H, D-18 > E] Rev-4FAc-FAEKFKEFVKDYFAKHWE-NH₂ 494

Examples of class A 4F and Rev 4F analogs with beta-Nph. Similarly,alpha-Nph analogs can be designed. Similarly to the above analogs, Hiscan be incorporated to Nph analogs. D>E analogs, E>D analogs and D-Eswitch analogs are additional possibilities similarly to the abovedescribed analogs.

4Nph Ac-DW Nph KA Nph YDKVAEK Nph KEA Nph -NH2 495 [D-E switched] Ac- EW Nph KA Nph Y E KVA D K Nph K D A Nph -NH2 496 4Nph [D > E]4Nph Ac- E WNph KA Nph Y E KVAEK Nph KEA Nph -NH2 497 [E > D]4Nph Ac-DW Nph KA NphYDKVA D K Nph K D A Nph -NH2 498 [D > E]4Nph Ac- E W Nph KA Nph YDKVAEKNph KEA Nph -NH2 499 [D > E]4Nph Ac-DW Nph KA Nph YEKVAEK Nph KEA Nph-NH2 500 [E-12 > D]4Nph Ac-DW Nph KA Nph YDKVA D K Nph KEA Nph -NH2 501[E-16 > D]4Nph Ac-DW Nph KA Nph YDKVAEK Nph K D A Nph -NH2 502

As described above for 4Nph, a minimum of 7 additional analogs for eachof the analogs given below.

[F-3, 6, > Nph] Ac-DW Nph KA Nph YDKVAEKFKEAF-NH2 503 4F [F-14, 18 >Nph] Ac-DWFKAFYDKVAEK Nph KEA Nph -NH2 504 4F [[F-3 > Nph] 4F Ac-DW NphKAFYDKVAEKFKEAF-NH2 505 [F-6 > Nph]4F Ac-DWFKA Nph YDKVAEKFKEAF-NH2 506[F-14 > Nph]4F Ac-DWFKAFYDKVAEK Nph KEAF-NH2 507 [F-18 > Nph]4FAc-DWFKAFYDKVAEKFKEA Nph -NH2 508

For each of the analog described below, a minimum of 7 additionalanalogs are possible as described above by switching D-E, D>E and E>Dand single D or E analogs.

Rev-4Nph Ac- Nph AEK Nph KEAVKDY Nph AK Nph 509 WD-NH2 [F-3, 6 > Nph]RevAc- Nph AEK Nph KEAVKDYFAKFWD-NH2 510 4F[F-13, 16]Rev- Ac-FAEKFKEAVKDYNph AK Nph WD-NH2 511 4F [F-3 > Nph]Rev-4F Ac- Nph AEKFKEAVKDYFAKFWD-NH2512 [F-6 > Nph]Rev-4F Ac-FAEK Nph KEAVKDYFAKFWD-NH2 513 [F-13 > Nph]Rev-Ac-FAEKFKEAVKDY Nph AKFWD-NH2 514 4F [F-16 > Nph]Rev- Ac-FAEKEKEAVKDYFAKNph WD-NH2 515 4F

For the analogs described below, additional analogs are possible byincorporating His or alpha-Nph and beta-Nph

Rev-[D > E]-4F Ac-FAEKFKEAVK E YFAKFW E -NH2 516 Rev-[E > D]4F Ac-FA DKFK D AVKDYFAKFWD-NH2 517 Rev-R4-4F Ac-FAE R FREAVKDYFAKFWD-NH2 518Rev-R6-4F Ac-FAEKF R EAVKDYFAKFWD-NH2 519 Rev-R10-4F Ac-FAEKFKEAV RDYFAKFWD-NH2 520 Rev-R14-4F Ac-FAEKFKEAVKDYFA R FWD-NH2 521 Rev-> E]-4FAc-FAEKFKEAVK E YFAKFW E -NH2 522 Rev-[E > D]4F Ac-FA D KFK DAVKDYFAKFWD-NH2 523 Rev-R4-4F Ac-FAE R FREAVKDYFAKFWD-NH2 524 Rev-R6-4FAc-FAEKF R EAVKDYFAKFWD-NH2 525 Rev-R10-4F Ac-FAEKFKEAV R DYFAKFWD-NH2526 Rev-R14-4F Ac-FAEKFKEAVKDYFA R FWD-NH2 527 Rev- > E]-4FAc-FAEKFKEAVK E YFAKFW E -NH2 528 Rev-[E > ]4F Ac-FA D KFK DAVKDYFAKFWD-NH2 529 Rev-R4-4F Ac-FAE R FREAVKDYFAKFWD-NH2 530 Rev-R6-4FAc-FAEKF R EAVKDYFAKFWD-NH2 531 Rev-R10-4F Ac-FAEKFKEAV R DYFAKFWD-NH2532 Rev-R14-4F Ac-FAEKFKEAVKDYFA R FWD-NH2 533 Rev-R4-4F Ac-FAE RFREAVKDYFAKFWD-NH2 534 Rev-R6-4F Ac-FAEKF R EAVKDYEAKFWD-NH2 535Rev-R10-4F Ac-FAEKFKEAV R DYFAKFWD-NH2 536 Rev-R14-4F Ac-FAEKFKEAVKDYFAR FWD-NH2 537 Rev-[D > E]-4F Ac-FAEKFKEAVK E YFAKFW E -NH2 538 Rev-[E >D]4F Ac-FA D KFK D AVKDYFAKFWD-NH2 539 Rev-R4-4F Ac-FAE RFREAVKDYFAKFWD-NH2 540 Rev-R6-4F Ac-FAEKF R EAVKDYFAKFWD-NH2 541Rev-R10-4F Ac-FAEKFKEAV R DYFAKFWD-NH2 542 Rev-R14-4F Ac-FAEKFKEAVKDYFAR FWD-NH2 543

For each of the analogs below, additional H and Nph analogs are possibleusing the examples described above. Each analog can yield 7 analogs withthe changes described in the examples given above.

Rev3F-2 Ac-LFEKFAEAFKDYVAKWKD-NH2 544 RevR4-3F-2 Ac-LFE RFAEAFKDYVAKWKD-NH2 545 RevR10-3F2 Ac-LFEKFAEAF R DYVAKWKD-NH2 546RevR15-3F-2 Ac-LFEKFAEAFKDYVA R WKD-NH2 547 RevR17-3F-2Ac-LFEKFAEAFKDYVAKW R D-NH2 548 Rev[D > E]3F2 Ac-LFEKFAEAFK E YVAKWK E-NH2 549 Rev[E] > D]3F-2 Ac-LF D KFA D AFKDYVAKWKD-NH2 550 Rev-[E3 >D]-3F-2 Ac-LF D KFAEAFKDYVAKWKD-NH2 551 Rev-[E7 > D]-3F-2 Ac-LFEKFA DAFKDYVAKWKD-NH2 552 Rev[D11 > E]3F-2 Ac-LFEKFAEAFK E YVAKWKD-NH2 553Rev-[D18 > E]3F-2 Ac-LFEKFAEAFKDYVAKWK E -NH2 554 Rev3F-1Ac-FAEKAWEFVKDYFAKLKD-NH2 555 RevR4-3F-1 Ac-FAE R AWEFVKDYFAKLKD-NH2 556RevR10-3F-1 Ac-FAEKAWEFV K DYFAKLKD-NH2 557 RevR15-3F-1Ac-FAEKAWEFVKDYFA K LKD-NH2 558 RevR17-3F-1 Ac-FAEKAWEFVKDYFAKL R D-NH2559 Rev[D > E]3F-1 Ac-FAEKAWEFVK E YFAKLKE-NH2 560 Rev[E > D}3F-1 Ac-FAD KAW D FVKDYFAKLKD-NH2 561 Rev[E3 > D]-3F-1 Ac-FA D KAWEFVKDYFAKLKD-NH2562 Rev[E7 > D]3F-1 Ac-FAEKAW D FVKDYFAKLKD-NH2 563 Rev-[D11 > E]3F-1Ac-FAEKAWEFVK E YFAKLKD-NH2 564 Rev-[D18 > E]3F-1 Ac-FAEKAWEFVKDYFAKLK E-NH2 565 Rev-5F Ac-FFEKFK E FVKDYFAKLWD-NH2 566 Rev-D > E]5FAc-FFEKFKEFVK E YFAKLW E -NH2 567 Rev-[E > D]5F Ac-FF D KFK DFVKDYFAKLWD-NH2 568 Rev-R4-5F Ac-FFE R FKEFVKDYFAKLWD-NH2 569 Rev-R6-5FAc-FFEKF R EFVKDYFAKLWD-NH2 570 Rev-R10-5F Ac-FFEKFKEFV R DYFAKLWD-NH2571 Rev-R15-5F Ac-FFEKFKEFVKDYFA R LWD-NH2 572 Rev-[E3 > D]-5F Ac-FF DKFKEFVKDYFAKLWD-NH2 573 Rev-[E7 > D]5F Ac-FFEKFK D FVKDYFAKLWD-NH2 574Rev-[D11 > E]-5F Ac-FFEKFKEFVK E YFAKLWD-NH2 575 Rev-[D18 > E]-5FAc-FFEKFKEFVKDYFAKLW E -NH2 576 Rev-5F-2 Ac-F L EKFKEFVKDYFAK F WD-NH2577 Rev-[D > E]-5F-2 Ac-FLEKFKEFVK E YFAKFW E -NH2 578 Rev-[E > D]-5F-2Ac-FL D KFK E FVKDYFAKFWD-NH2 579 Rev-[E3 > D]-5F-2 Ac-FL DKFKEFVKDYFAKFWD-NH2 580 Rev-[E7 > D]-5F-2 Ac-FLEKFK D FVKDYFAKFWD-NH2581 Rev-[D11 > E]-5F-2 Ac-FLEKFKEFVK E YFAKFWD-NH2 582 Rev-[D18 >E]-5F-2 Ac-FLEKFKEFVKDYFAKFW E -NH2 583 Rev-R4-5F-2 Ac-FLE RFKEFVKDYFAKFWD-NH2 584 Rev-R6-5F-2 Ac-FLEKF R EFVKDYFAKFWD-NH2 585RevR10-5F-2 Ac-FLEKFKEFV R DYFAKFWD-NH2 586 Rev-R16-5F-2Ac-FLEKFKEFVKDYFA R FWD-NH2 587 Rev-6F Ac-F F EK F KE FF KDYFAKLWD-NH2588 Rev-[D > E]-6F Ac-FFEKFKEFFK E YFAKLW E -NH2 589 Rev-[E > D]-6FAc-FF D KFK D FFKDYFAKLWD-NH2 590 Rev-R4-6F Ac-FFE R FKEFFKDYFAKLWD-NH2591 Rev-R6-6F Ac-F F EKF R EFFKDYFAKLWD-NH2 592 Rev-R10-6F Ac-FFEKYKEFFR DYFAKLWD-NH2 593 Rev-R14-6F Ac-FFERFKEFFKDYFA R LWD-NH2 594 Rev-[E3 >D]-6F Ac-FF D KFKEFFKDYFAKLWD-NH 595 Rev-[E7 > D]-6F Ac-FFEKEK DFFKDYFAKLWD-NH2 596 Rev-[D11 > E]-6F Ac-FFEKFKEFFK E YFAKLWD-NH2 597Rev-[D18 > E]-6F  Ac-FFEKFKEFFKDYFAKLW E -NH2 598 Rev-4FAc-FAEKFKEAVKDYFAKFWD-NH2 599 Rev-[D > E]-4F Ac-FAEKFKEAVK E YFAKFW E-NH2 600 Rev-[E > D]4F Ac-FA D KFK D AVKDYFAKFWD-NH2 601 Rev-R4-4FAc-FAE R FREAVKDYFAKFWD-NH2 602 Rev-R6-4F Ac-FAEKF R EAVKDYFAKFWD-NH2603 Rev-R10-4F Ac-FAEKFKEAV R DYFAKFWD-NH2 604 Rev-R14-4FAc-FAEKFKEAVKDYFA R FWD-NH2 605 4F-2 Ac-DKWKAVYDKFAEAFKEFF-NH2 606 [D >E]-4F-2 Ac-EKWKAVYEKFAEAFKEFF-NH2 607 [E > D]-4F-2 Ac-DKWKAVYDKFA D AFKD FF-NH2 608 R2-4F-2 Ac-D R WKAVYDKFAEAFKEFF-NH2 609 R4-4F-2 Ac-DKW RAVYDKFAEAFKEFF-NH2 610 R9-4F-2 Ac-DKWKAVYD R FAEAFKEFF-NH2 611 R14-4F-2Ac-DKWKAVYDKFAEAF R EFF-NH2 612 Rev4F-2 Ac-FFEKFAEAFKDYVAKWKD-NH2 613Rev-[D > E]-4F-2 Ac-FFEKFAEAFK E YVAKWK E -NH2 614 Rev-[E > D]-3F-2Ac-FF D KFA D AFKDYVAKWKD-NH2 615 Rev-R4-4F-2 Ac-FFE RFAEAFKDYVAKWKD-NH2 616 Rev-R10-4F-2 Ac-EFERFAEAF R DYVAKWKD-NH2 617Rev-R15-4F-2 Ac-FFEKFAEAFKDYVA R WKD-NH2 618 Rev-R17-4F-2 Ac-FFE RFAEAFKDYVAKW R D-NH2 619 Rev-[E3 > D]-4F-2  Ac-FF D KFAEAFKDYVAKWKD-NH2620 Rev-[E7 > D] -4F-2 Ac-FFEKFA D AFKDYYAKWKD-NH2 621 Rev-[D11 >E]-4F-2 Ac-FFERFAEAFK E YVAKWKD-NH2 622 Rev-[D18 > E]-4F-2Ac-FFERFAEAFKDYVAKWKE-NH2 623 Rev-7F Ac-FFEKFKEFFKDYFAKFWD-NH2 624Rev-[E > D]-7F Ac-FF D KFK D FFKDYFAKFWD-NH2 625 Rev-[D > E]-7FAc-FFEKFKEFFK E YFAKFW E -NH2 626 Rev-R4-7F Ac-FFE R FKEFFKDYFAKFWD-NH2627 Rev-R6-7F Ac-FFEKF R EFFKDYFAKFWD-NH2 628 Rev-R10-7F Ac-FFEKFKEFF RDYFAKFWD-NH2 629 Rev-R14-7F Ac-FFEKFKEFFKDYFA R FWD-NH2 630 Rev-[E3 >D]-7F Ac-FF D KFKEFFKDYFAKFWD-NH2 631 Rev-[E7 > D]7F Ac-FFEKFK DFFKDYFAKFWD-NH2 632 Rev-[D11 >E]-7F Ac-FFEKFKEFFK E YFAKFWD-NH2 633Rev-[D18 > E]-7F Ac-FFEKFKEFFKDYFAKFW E -NH2 634

It is also noted that any of the peptides described herein can comprisenon-natural amino acids in addition to or instead of the correspondingthe natural amino acids identified herein. Such modifications include,but are not limited to acetylation, amidation, formylation, methylation,sulfation, and the like. Illustrative non-natural amino acids include,but are not limited to Ornithine, norleucine, norvaline, N-methylvaline,6-N-methyllysine, N-methylisoleucine, N-methylglycine, sarcosine,inosine, allo-isoleucine, isodesmolysine, 4-hydroxyproline,3-hydroxyproline, allo-hydroxylysine, hydroxylisine, N-ethylasparagine,N-ethylglycine, 2,3-diaminopropionic acid, 2,2′-diaminopropionic acid,desmosine, 2,4-diaminobutyric acid, 2-aminopimelic acid,3-aminoisobutyric acid, 2-aminoisobutyric acid, 2-aminoheptanoic acid,6-aminocaproic acid, 4-aminobutyric acid, 2-aminobutyric acid,beta-alanine, 3-aminoadipic acid, 2-aminoadipic acid, and the like. Incertain embodiments and one or more of the “natural” amino acids of thepeptides described herein, can be substituted with the correspondingnon-natural amino acid (e.g. as describe above).

In certain embodiments, this invention contemplates particularly the useof modified lysines. Such modifications include, but are not limited to,biotin modification of epsilon lysines and/or methylation of the epsilonlysines. Illustrative peptide comprising epsilon methylated lysinesinclude, but are not limited to:Ac-D-W-F-K(eCH₃)₂-A-F-Y-D-K(eCH₃)₂-V-A-E-K(eCH₃)-₂-F-K(eCH₃)₂-E-A-F-NH(CH₃)₂(SEQ ID NO:635) and:Ac-DWFK(eCH₃)₂AFYDK(eCH₃)₂VAEK(eCH₃)₂FK(eCH₃)₂EAF-NH(CH₃) (SEQ IDNO:636). Other modified amino acids include but are not limited toornithine analogs and homoaminoalanine analogs (instead of (CH₂)₄—NH₂for Lys it can be —(CH₂)₂—NH₂ for Haa and —(CH₂)₃—NH₂ for Orn] and thelike. It is noted that these modifications are illustrative and notintended to be limiting. Illustrative 4F analogues that possess modifiedamino acids are shown in Table 6.

TABLE 6 Illustrative 4F analogs that comprise modified amino acids. εN-Dimethyl-Lys derivative of 4F (εN-Dime)Ac-D-W-F-K(εN-Dime)-A-F-Y-D-K(εN-Dime)-V- 637A-E-K(εN-Dime)-F-K(εN-Dime)-E-A-F-NH₂Ac-D-W-F-K-(εN-Dime)-A-F-Y-D-K(εN-Dime)-V- 638A-E-K(εN-Dime)-F-K((εN-Dime)-E-A-F-NH-MeAc-D-W-F-K-(εN-Dime)-A-F-Y-D-K(εN-Dime)-V- 639A-E-K(εN-Dime)-F-K(εN-Dime)-E-A-F-N-(Me)₂εN-Diethyl-Lys derivatives of 4F (εN-Diet)Ac-D-W-F-K(εN-Diet)-A-F-Y-D-K(εN-Diet)-V- 640A-E-K(εN-Diet)-F-K(εN-Diet)-E-A-F-NH₂Ac-D-W-F-K(εN-Diet)-A-F-Y-D-K(εN-Diet)-V- 641A-E-K(εN-Diet)-F-K(εN-Diet)-E-A-F-NH-EtAc-D-W-F-K(εN-Diet)-A-F-Y-D-K(εN-Diet)-V- 642A-E-K(εN-Diet)-F-K(εN-Diet)-E-A-F-NH-(Et)₂εN-Monomethyl-Lys derivative of 4F (εN-Me)Ac-D-W-F-K(εN-Me)-A-F-Y-D-K(εN-Me)-V-A- 643E-K(εN-Me)-F-K(εN-Me)-E-A-F-NH₂ Ac-D-W-F-K(εN-Me)-A-F-Y-D-K(εN-Me)-V-A-644 E-K(εN-Me)-F-K(εN-Me)-E-A-F-NH-MeAc-D-W-F-K(εN-Me)-A-F-Y-D-K(εN-Me)-V-A- 645E-K(εN-Me)-F-K(εN-Me)-E-A-F-N-(Me)₂ εN-ethylLys derivative of 4F (εN-Et)Ac-D-W-F-K(εN-Et)-A-F-Y-D-K(εN-E0-V-A- 646E-K(εN-Et)-F-K(εN-Et)-E-A-F-NH₂ Ac-D-W-F-K(εN-Et)-A-F-Y-D-K(εN-E0-V-A-647 E-K(εN-Et)-F-K(εN-E0-E-A-F-NH-EtAc-D-W-F-K(εN-Et)-A-F-Y-D-K(εN-Et)-V-A- 648E-K(εN-Et)-F-K(εN-Et)-E-A-F-NH-(Et)₂HomoLys analogs of 4F(hK)(--CH₂)_(ζ5)-NH₂Ac-D-W-F-hK-A-F-Y-D-hK-V-A-E-hK-F-hK-E-A-F-NH₂ 649Ac-D-W-F-hK(εN-Dime)-A-F-Y-D-hK(εN-Dime)-V- 650A-E-hK(εN-Dime)-F-hK(εN-Dime)-E-A-F-NH₂Ac-D-W-F-hK(εN-Dime)-A-F-Y-D-hK(εN-Dime)-V- 651A-E-hK(εN-Dime)-F-hK(εN-Dime)-E-A-F-N-(Me)₂Ac-D-W-F-hK(εN-Dime)-A-F-Y-D-hK(εN-Dime)-V- 652A-E-hK(εN -Dime)-F-hK(εN-Dime)-E-A-F-NH-MeAc-D-W-F-hK(εN-Diet)-A-F-Y-D-hK(εN-Diet)-V- 653A-E-hK(εN -Diet)-F-hK(N-Diet)-E-A-F-NH-EtAc-D-W-F-hK(εN-Me)-A-F-Y-D-hK(εN-Me)-V- 654A-E-hK(εN-Me)-F-hK(εN-Me)-E-A-F-NHAc-D-W-F-hK(εN-Me)-A-F-Y-D-hK(εN-Me)-V- 655A-E-hK(εN-Me)-F-hK(εN-Me)-E-A-F-NH-MeAc-D-W-F-hK(εN-Me)-A-F-Y-D-hK(εN-Me)-V- 656A-E-hK(εN-Me)-F-hK(εN-Me)-E-A-F-N-(Me)₂Ac-D-W-F-hK(εN-Et)-A-F-Y-D-hK(εN-Et)-V- 657A-E-hK(εN-Et)-F-hK(εN-Et)-E-A-F-NH₂Ac-D-W-F-hK(εN-Et-A-F-Y-D-hK(εN-Et)-V- 658A-E-hK(εN-Et)-F-hK(N-Et)-E-A-F-NH-EtAc-D-W-F-hK(εN-Et)-A-F-Y-D-hK(εN-Et)-V- 659A-E-hK(N-Et)-F-hK(εN-Et)-E-A-F-NH-(Et)₂4F analogs in which K is replaced O 660 (O = Ornithine, --(CH₂)₃-NH₂)Ac-D-W-F-O-A-F-Y-D-O-V-A-E-O-F-O-E-A-F-NH₂ 661Ac-D-W-F-O(δ-Dime)-A-F--Y-D-O(δN-Dime)- 662V-A-E-O(δN-Dime)-F-O(δN-Dime)-E-A-F-NH₂Ac-D-W-F-O(δN-Dime)-A-F-Y-D-)(δN-Dime)- 663V-A-E-O(δN-Dime)-F-O(δN-Dime)-E-A-F-N-(Me)₂Ac-D-W-F-O(δN-Dime)-A-F-Y-D-O(δ3N-Dime)- 664V-A-E-O(δN-Dime)-F-O(δ5N-Dime)-E-A-F-NH-MeAc-D-W-F-O(δN-Diet)-A-F-Y-D-O(δN-Diet)- 665V-A-E-O(δN-Diet)-F-O(δN-Diet)-E-A-F-NH-EtAc-D-W-F-O(δN-Me)-A-F-Y-D-O(δN-Me)-V-A-E-O 666(δN-Me)-F-O(δN-Me)-E-A-F-NH₂ Ac-D-W-F-O(δN-Me)-A-F-Y-D-O(δN-Me)-V-A-E-O667 (δN-Me)-F-O(δ3N-Me)-E-A-F-NH-MeAc-D-W-F-O(δN-Me)-A-F-Y-D-O(δN-Me)-V-A-E-O 668(δN-Me)-F-O(δN-Me)-E-A-F-N-(Me)₂Ac-D-W-F-O(δN-Et)-A-F-Y-D-O(δN-Et)-V-A-E-O 669(δN-Et)-F-O(δN-Et)-E-A-F-NH₂ Ac-D-W-F-O(δN-ED-A-F-Y-D-O(δN-Et)-V-A-E-O670 (δN-Et)-F-O(δN-Et)-E-A-F-NH-EtAc-D-W-F-O(δN-Et)-A-F-Y-D-O(δN-Et)-V-A-E- 671O(δN-Et)-F-O(δN-Et)-E-A-F-NH-(Et)₂

The peptides and modifications shown above are intended to beillustrative and not limiting.

E) Smaller Peptides.

It was also a surprising discovery that certain small peptidesconsisting of a minimum of three amino acids preferentially (but notnecessarily) with one or more of the amino acids being theD-stereoisomer of the amino acid, and possessing hydrophobic domains topermit lipid protein interactions, and hydrophilic domains to permit adegree of water solubility also possess significant anti-inflammatoryproperties and are useful in treating one or more of the pathologiesdescribed herein. The “small peptides” typically range in length from 2amino acids to about 15 amino acids, more preferably from about 3 aminoacids to about 10 or 11 amino acids, and most preferably from about 4 toabout 8 or 10 amino acids. In various embodiments the peptides aretypically characterized by having hydrophobic terminal amino acids orterminal amino acids rendered hydrophobic by the attachment of one ormore hydrophobic “protecting” groups. Various “small peptides” aredescribed in copending applications U.S. Ser. No. 10/649,378, filed Aug.26, 2003, and in U.S. Ser. No. 10/913,800, filed on Aug. 6, 2004, and inPCT Application PCT/US2004/026288.

In certain embodiments, the peptides can be characterized by Formula I,below: X¹-X²-X³ _(n)-X⁴I where, n is 0 or 1, X¹ is a hydrophobic aminoacid and/or bears a hydrophobic protecting group, X⁴ is a hydrophobicamino acid and/or bears a hydrophobic protecting group; and when n is 0X² is an acidic or a basic amino acid; when n is 1: X² and X³ areindependently an acidic amino acid, a basic amino acid, an aliphaticamino acid, or an aromatic amino acid such that when X² is an acidicamino acid; X³ is a basic amino acid, an aliphatic amino acid, or anaromatic amino acid; when X² is a basic amino acid; X³ is an acidicamino acid, an aliphatic amino acid, or an aromatic amino acid; and whenX² is an aliphatic or aromatic amino acid, X³ is an acidic amino acid,or a basic amino acid.

Longer peptides (e.g., up to 10, 11, or 15 amino acids) are alsocontemplated within the scope of this invention. Typically where theshorter peptides (e.g., peptides according to formula I) arecharacterized by an acidic, basic, aliphatic, or aromatic amino acid,the longer peptides are characterized by acidic, basic, aliphatic, oraromatic domains comprising two or more amino acids of that type.

1) Functional Properties of Active Small Peptides.

It was a surprising finding of this invention that a number of physicalproperties predict the ability of small peptides (e.g., less than 10amino acids, preferably less than 8 amino acids, more preferably fromabout 3 to about 5 or 6 amino acids) of this invention to render HDLmore anti-inflammatory and to mitigate atherosclerosis and/or otherpathologies characterized by an inflammatory response in a mammal. Thephysical properties include high solubility in ethyl acetate (e.g.,greater than about 4 mg/mL), and solubility in aqueous buffer at pH 7.0.Upon contacting phospholipids such as1,2-Dimyristoyl-sn-glycero-3-phosphocholine (DMPC), in an aqueousenvironment, the particularly effective small peptides induce orparticipate in the formation of particles with a diameter ofapproximately 7.5 nm (±0.1 nm), and/or induce or participate in theformation of stacked bilayers with a bilayer dimension on the order of3.4 to 4.1 nm with spacing between the bilayers in the stack ofapproximately 2 nm, and/or also induce or participate in the formationof vesicular structures of approximately 38 nm). In certain preferredembodiments, the small peptides have a molecular weight of less thanabout 900 Da.

Thus, in certain embodiments, this invention contemplates small peptidesthat ameliorate one or more symptoms of an indication/pathologydescribed herein, e.g., an inflammatory condition, where the peptide(s):ranges in length from about 3 to about 8 amino acids, preferably fromabout 3 to about 6, or 7 amino acids, and more preferably from about 3to about 5 amino acids; are soluble in ethyl acetate at a concentrationgreater than about 4 mg/mL; are soluble in aqueous buffer at pH 7.0;when contacted with a phospholipid in an aqueous environment, formparticles with a diameter of approximately 7.5 nm and/or form stackedbilayers with a bilayer dimension on the order of 3.4 to 4.1 nm withspacing between the bilayers in the stack of approximately 2 nm; have amolecular weight less than about 900 daltons; convert pro-inflammatoryHDL to anti-inflammatory HDL or make anti-inflammatory HDL moreanti-inflammatory; and do not have the amino acid sequenceLys-Arg-Asp-Ser (SEQ ID NO:801), especially in which Lys-Arg-Asp and Serare all L amino acids. In certain embodiments, these small peptidesprotect a phospholipid against oxidation by an oxidizing agent.

While these small peptides need not be so limited, in certainembodiments, these small peptides can include the small peptidesdescribed below.

2) Tripeptides.

It was discovered that certain tripeptides (3 amino acid peptides) canbe synthesized that show desirable properties as described herein (e.g.,the ability to convert pro-inflammatory HDL to anti-inflammatory HDL,the ability to decrease LDL-induced monocyte chemotactic activitygenerated by artery wall cells, the ability to increase pre-beta HDL,etc.). In certain embodiments, the peptides are characterized by formulaI, wherein N is zero, shown below as Formula II: X¹-X²-X⁴ II where theend amino acids (X¹ and X⁴) are hydrophobic either because of ahydrophobic side chain or because the side chain or the C and/or Nterminus is blocked with one or more hydrophobic protecting group(s)(e.g., the N-terminus is blocked with Boc-, Fmoc-, nicotinyl-, etc., andthe C-terminus blocked with (tBu)-OtBu, etc.). In certain embodiments,the X² amino acid is either acidic (e.g., aspartic acid, glutamic acid,etc.) or basic (e.g., histidine, arginine, lysine, etc.). The peptidecan be all L-amino acids or include one or more or all D-amino acids.

Certain preferred tripeptides of this invention include, but are notlimited to the peptides shown in Table 7.

TABLE 7 Examples of certain preferred tripeptides bearing hydrophobic blocking groups and acidic, basic, or histidine central amino acids. SEQ ID X¹ X² X³ X⁴ NOBoc-Lys(EBoc) Arg Ser(tBu)-OtBu 672 Boc-Lys(EBoc) Arg Thr(tBu)-OtBu 673Boc-Trp Arg Ile-OtBu 674 Boc-Trp Arg Leu-OtBu 675 Boc-Phe Arg Ile -OtBu676 Boc-Phe Arg Leu-OtBu 677 Boc-Lys(EBoc) Glu Ser(tBu)-OtBu 678Boc-Lys(EBoc) Glu Thr(tBu)-OtBu 679 Boc-Lys(EBoc) Asp Ser(tBu)-OtBu 680Boc-Lys(EBoc) Asp Thr(tBu)-OtBu 681 Boc-Lys(EBoc) Arg Ser(tBu)-OtBu 682Boc-Lys(EBoc) Arg Thr(tBu)-OtBu 683 Boc-Leu Glu Ser(tBu)-OtBu 684Boc-Leu Glu Thr(tBu)-OtBu 685 Fmoc-Trp Arg Ser(tBu)-OtBu 686 Fmoc-TrpAsp Ser(tBu)-OtBu 687 Fmoc -Trp Glu Ser(tBu)-OtBu 688 Fmoc-Trp ArgSer(tBu)-OtBu 689 Boc-Lys(EBoc) Glu Leu-OtBu 690 Fmoc-Leu ArgSer(tBu)-OtBu 691 Fmoc-Leu Asp Ser(tBu)-OtBu 692 Fmoc-Leu GluSer(tBu)-OtBu 693 Fmoc-Leu Arg Ser(tBu)-OtBu 694 Fmoc-Leu ArgThr(tBu)-OtBu 695 Boc-Glu Asp Tyr(tBu)-OtBu 696 Fmoc-Lys(eFmoc) ArgSer(tBu)-OtBu 697 Fmoc-Trp Arg Ile-OtBu 698 Fmoc-Trp Arg Leu-OtBu 699Fmoc-Phe Arg Ile-OtBu 700 Fmoc-Phe Arg Leu-OtBu 701 Boc-Trp Arg Phe-OtBu702 Boc-Trp Arg Tyr-OtBu 703 Fmoc-Trp Arg Phe-OtBu 704 Fmoc-Trp ArgTyr-OtBu 705 Boc-Orn(OBoc) Arg Ser(tBu)-OtBu 706 Nicotinyl ArgSer(tBu)-OtBu 707 Lys(sBoc) Nicotinyl Arg Thr(tBu)-OtBu 708 Lys(EBoc)Fmoc-Leu Asp Thr(tBu)-OtBu 709 Fmoc-Leu Glu Thr(tBu)-OtBu 710 Fmoc-LeuArg Thr(tBu)-OtBu 711 Fmoc-norLeu Arg Ser(tBu)-OtBu 712 Fmoc-norLeu AspSer(tBu)-OtBu 713 Fmoc-norLeu Glu Ser(tBu)-OtBu 714 Fmoc-Lys(cBoc) ArgSer(tBu)-OtBu 715 Fmoc-Lys(EBoc) Arg Thr(tBu)-OtBu 716 Fmoc-Lys(EBoc)Glu Ser(tBu)-OtBu 717 Fmoc-Lys(EBoc) Glu Thr(tBu)-OtBu 718Fmoc-Lys(cBoc) Asp Ser(tBu)-OtBu 719 Fmoc-Lys(EBoc) Asp Thr(tBu)-OtBu720 Fmoc-Lys(eBoc) Glu Leu-OtBu 721 Fmoc-Lys(EBoc) Arg Leu-OtBu 722Fmoc-Lys(cFmoc) Arg Thr(tBu)-OtBu 723 Fmoc- Lys(EFmoc) Glu Ser(tBu)-OtBu724 Fmoc- Lys(EFmoc) Glu Thr(tBu)-OtBu 725 Fmoc- Lys(cFmoc) AspSer(tBu)-OtBu 726 Fmoc- Lys(EFmoc) Asp Thr(tBu)-OtBu 727Fmoc- Lys(EFmoc) Arg Ser(tBu)-OtBu 728 Fmoc- Glu Leu-OtBu 729 Lys(EFmoc)Boc-Lys(EFmoc) Asp Ser(tBu)-OtBu 730 Boc-Lys(EFmoc) Asp Thr(tBu)-OtBu731 Boc-Lys(EFmoc) Arg Thr(tBu)-OtBu 732 Boc-Lys(EFmoc) Glu Leu-OtBu 733Boc-Orn(8Fmoc) Glu Ser(tBu)-OtBu 734 Boc-Orn(8Fmoc) Asp Ser(tBu)-OtBu735 Boc-Orn(6Fmoc) Asp Thr(tBu)-OtBu 736 Boc-Orn(OFmoc) ArgThr(tBu)-OtBu 737 Boc-Orn(6Fmoc) Glu Thr(tBu)-OtBu 738 Fmoc-Trp AspIle-OtBu 739 Fmoc-Trp Arg Ile-OtBu 740 Fmoc-Trp Glu Ile-OtBu 741Fmoc-Trp Asp Leu-OtBu 742 Fmoc-Trp Glu Leu-OtBu 743 Fmoc-Phe AspIle-OtBu 744 Fmoc-Phe Asp Leu-OtBu 745 Fmoc-Phe Glu Leu-OtBu 746Fmoc-Trp Arg Phe-OtBu 747 Fmoc-Trp Glu Phe-OtBu 748 Fmoc-Trp AspPhe-OtBu 749 Fmoc-Trp Asp Tyr-OtBu 750 Fmoc-Trp Arg Tyr-OtBu 751Fmoc-Trp Glu Tyr-OtBu 752 Fmoc-Trp Arg Thr(tBu)-OtBu 753 Fmoc-Trp AspThr(tBu)-OtBu 754 Fmoc-Trp Glu Thr(tBu)-OtBu 755 Boc-Phe Arg norLeu-OtBu756 Boc-Phe Glu norLeu-OtBu 757 Fmoc-Phe Asp norLeu-OtBu 758 Boc-Glu HisTyr(tBu)-OtBu 759 Boc-Leu His Ser(tBu)-OtBu 760 Boc-Leu HisThr(tBu)-OtBu 761 Boc-Lys(EBoc) His Ser(tBu)-OtBu 762 Boc-Lys(EBoc) HisThr(tBu)-OtBu 763 Boc-Lys(EBoc) His Leu-OtBu 764 Boc-Lys(EFmoc) HisSer(tBu)-OtBu 765 Boc-Lys(eFmoc) His Thr(tBu)-OtBu 766 Boc-Lys(EFmoc)His Leu-OtBu 767 Boc-Orn(OBoc) His Ser(tBu)-OtBu 768 Boc-Orn(6Fmoc) HisThr(tBu)-OtBu 769 Boc-Phe His De-OtBu 770 Boc-Phe His Leu-OtBu 771Boc-Phe His norLeu-OtBu 772 Boc-Phe Lys Leu-OtB u 773 Boc-Trp HisIle-OtBu 774 Boc-Trp His Leu-OtBu 775 Boc-Trp His Phe-OtBu 776 Boc-TrpHis Tyr-OtBu 777 Boc-Phe Lys Leu-OtBu 778 Fmoc-Lys(EFmoc) HisSer(tBu)-OtBu 779 Fmoc-Lys(EFmoc) His Thr(tBu)-OtBu 780 Fmoc-Lys(EFmoc)His Leu-OtBu 781 Fmoc-Leu His Ser(tBu)-OtBu 782 Fmoc-Leu HisThr(tBu)-OtBu 783 Fmoc-Lys(EBoc) His Ser(tB u)-OtBu 784 Fmoc-Lys(EBoc)His Thr(tBu)-OtBu 785 Fmoc-Lys(eBoc) His Leu-OtBu 786 Fmoc-Lys(EFmoc)His Ser(tBu)-OtBu 787 Fmoc-Lys(EFmoc) His Thr(tBu)-OtBu 788 Fmoc-norLeuHis Ser(tBu)-OtBu 789 Fmoc-Phe His Ile-OtBu 790 Fmoc-Phe His Leu-OtBu791 Fmoc-Phe His norLeu-OtBu 792 Fmoc-Trp His Ser(tBu)-OtBu 793 Fmoc-TrpHis Ile-OtBu 794 Fmoc-Trp His Leu-OtBu 795 Fmoc-Trp His Phe-OtBu 796Fmoc-Trp His Tyr-OtBu 797 Fmoc-Trp His Thr(tBu)-OtBu 798 Nicotinyl HisSer(tBu)-OtBu 799 Lys(eBoc) Nicotinyl His Thr(tBu)-OtBu 800 Lys(EBoc)

While the peptides of Table 7 are illustrated with particular protectinggroups, it is noted that these groups may be substituted with otherprotecting groups as described herein and/or one or more of the shownprotecting group can be eliminated.

3) Small Peptides with Central Acidic and Basic Amino Acids.

In certain embodiments, the peptides of this invention range from fouramino acids to about ten amino acids. The terminal amino acids aretypically hydrophobic either because of a hydrophobic side chain orbecause the terminal amino acids bear one or more hydrophobic protectinggroups end amino acids (X¹ and X⁴) are hydrophobic either because of ahydrophobic side chain or because the side chain or the C and/or Nterminus is blocked with one or more hydrophobic protecting group(s)(e.g., the N-terminus is blocked with Boc-, Fmoc-, Nicotinyl-, etc., andthe C-terminus blocked with (tBu)-OtBu, etc.). Typically, the centralportion of the peptide comprises a basic amino acid and an acidic aminoacid (e.g., in a 4 mer) or a basic domain and/or an acidic domain in alonger molecule.

These four-mers can be represented by Formula I in which X¹ and X⁴ arehydrophobic and/or bear hydrophobic protecting group(s) as describedherein and X² is acidic while X³ is basic or X² is basic while X³ isacidic. The peptide can be all L-amino acids or include one or more orall D-amino acids.

Certain preferred peptides of this invention include, but are notlimited to the peptides shown in Table 8.

TABLE 8 Illustrative examples of small peptideswith central acidic and basic amino acids. SEQ ID X¹ X² X³ X⁴ NOBoc-Lys(EBoc) Arg Asp Ser(tBu)-OtBu 801 Boc-Lys(EBoc) Arg AspThr(tBu)-OtBu 802 Boc-Trp Arg Asp Ile-OtBu 803 Boc-Trp Arg Asp Leu-OtBu804 Boc-Phe Arg Asp Leu-OtBu 805 Boc-Phe Arg Asp Ile-OtBu 806 Boc-PheArg Asp norLeu-OtBu 807 Boc-Phe Arg Glu norLeu-OtBu 808 Boc-Phe Arg GluIle-OtBu 809 Boc-Phe Asp Arg Ile-OtBu 810 Boc-Phe Glu Arg Ile-OtBu 811Boc-Phe Asp Arg Leu-OtBu 812 Boc-Phe Arg Glu Leu-OtBu 813 Boc-Phe GluArg Leu-OtBu 814 Boc-Phe Asp Arg norLeu-OtBu 815 Boc-Phe Glu ArgnorLeu-OtBu 816 Boc-Lys(EBoc) Glu Arg Ser(tBu)-OtBu 817 Boc-Lys(EBoc)Glu Arg Thr(tBu)-OtBu 818 Boc-Lys(EBoc) Asp Arg Ser(tBu)-OtBu 819Boc-Lys(EBoc) Asp Arg Thr(tBu)-OtBu 820 Boc-Lys(eBoc) Arg GluSer(tBu)-OtBu 821 Boc-Lys(eBoc) Arg Glu Thr(tBu)-OtBu 822 Boc-Leu GluArg Ser(tBu)-OtBu 823 Boc-Leu Glu Arg Thr(tBu)-OtBu 824 Fmoc-Trp Arg AspSer(tBu)-OtBu 825 Fmoc-Trp Asp Arg Ser(tBu)-OtBu 826 Fmoc-Trp Glu ArgSer(tBu)-OtBu 827 Fmoc-Trp Arg Glu Ser(tBu)-OtBu 828 Boc-Lys(eBoc) GluArg Leu-OtBu 829 Fmoc-Leu Arg Asp Ser(tBu)-OtBu 830 Fmoc-Leu Asp ArgSer(tBu)-OtBu 831 Fmoc-Leu Glu Arg Ser(tBu)-OtBu 832 Fmoc-Leu Arg GluSer(tBu)-OtBu 833 Fmoc-Leu Arg Asp Thr(tBu)-OtBu 834 Boc-Glu Asp ArgTyr(tBu)-OtBu 835 Fmoc-Lys(EFmoc) Arg Asp Ser(tBu)-OtBu 836 Fmoc-Trp ArgAsp Ile-OtBu 837 Fmoc-Trp Arg Asp Leu-OtBu 838 Fmoc-Phe Arg AspIle-OtB u 839 Fmoc-Phe Arg Asp Leu-OtBu 840 Boc-Trp Arg Asp Phe-OtBu 841Boc-Trp Arg Asp Tyr-OtBu 842 Fmoc-Trp Arg Asp Phe-OtBu 843 Fmoc-Trp ArgAsp Tyr-OtBu 844 Boc-Orn(8Boc) Arg Glu Ser(tBu)-OtBu 845 Nicotinyl ) ArgAsp Ser(tBu)-OtBu 846 Lys(eBoc Nicotinyl  Arg Asp Thr(tBu)-OtBu 847Lys(EBoc) Fmoc-Leu Asp Arg Thr(tBu)-OtBu 848 Fmoc-Leu Glu ArgThr(tBu)-OtBu 849 Fmoc-Leu Arg Glu Thr(tBu)-OtBu 850 Fmoc-norLeu Arg AspSer(tBu)-OtBu 851 Fmoc-norLeu Asp Arg Ser(tBu)-OtBu 852 Fmoc-norLeu GluArg Ser(tBu)-OtBu 853 Fmoc-norLeu Arg Glu Ser(tBu)-OtBu 854Fmoc-Lys(EBoc) Arg Asp Ser(tBu)-OtBu 855 Fmoc-Lys(EBoc) Arg AspThr(tBu)-OtBu 856 Fmoc-Lys(EBoc) Glu Arg Ser(tBu)-OtBu 857Fmoc-Lys(EBoc) Glu Arg Thr(tBu)-OtBu 858 Fmoc-Lys(eBoc) Asp ArgSer(tBu)-OtBu 859 Fmoc-Lys(EBoc) Asp Arg Thr(tBu)-OtBu 860Fmoc-Lys(EBoc) Arg Glu Ser(tBu)-OtBu 861 Fmoc-Lys(EBoc) Arg GluThr(tBu)-OtBu 862 Fmoc-Lys(EBoc) Glu Arg Leu-OtBu 863 Fmoc-Lys(EBoc) ArgGlu Leu-OtBu 864 Fmoc-Lys(EFmoc) Arg Asp Thr(tBu)-OtBu 865Fmoc-Lys(EFmoc) Glu Arg Ser(tBu)-OtBu 866 Fmoc-Lys(EFmoc) Glu ArgThr(tBu)-OtBu 867 Fmoc-Lys(EFmoc) Asp Arg Ser(tBu)-OtBu 868Fmoc-Lys(cFmoc) Asp Arg Thr(tBu)-OtBu 869 Fmoc-Lys(EFmoc) Arg GluSer(tBu)-OtBu 870 Fmoc-Lys(EFmoc) Arg Glu Thr(tBu)-OtBu 871Fmoc-Lys(EFmoc)) Glu Arg Leu-OtBu 872 Boc-Lys(eFmoc) Arg AspSer(tBu)-OtBu 873 Boc-Lys(EFmoc) Arg Asp Thr(tBu)-OtBu 874Boc-Lys(EFmoc) Glu Arg Ser(tBu)-OtBu 875 Boc-Lys(eFmoc) Glu ArgThr(tBu)-OtBu 876 Boc-Lys(EFmoc) Asp Arg Ser(tBu)-OtBu 877Boc-Lys(eFmoc) Asp Arg Thr(tBu)-OtBu 878 Boc-Lys(EFmoc) Arg GluSer(tBu)-OtBu 879 Boc-Lys(EFmoc) Arg Glu Thr(tBu)-OtBu 880Boc-Lys(EFmoc) Glu Arg Leu-OtBu 881 Boc-Om(OFmoc) Arg Glu Ser(tBu)-OtBu882 Boc-Om(8Fmoc) Glu Arg Ser(tBu)-OtBu 883 Boc-Orn(8Fmoc) Arg AspSer(tBu)-OtBu 884 Boc-Om(8Fmoc) Asp Arg Ser(tBu)-OtBu  885Boc-Orn(OFmoc)  Asp  Arg Thr(tBu)-OtBu 886 Boc-Om(8Fmoc) Arg AspThr(tBu)-OtBu  887 Boc-Orn(8Fmoc) Glu Arg Thr(tBu)-OtBu  888Boc-Orn(OFmoc)  Arg Glu  Thr(tBu)-OtBu 889 Fmoc-Trp Asp Arg Ile-OtBu 890Fmoc-Trp Arg Glu Ile-OtBu 891 Fmoc-Trp Glu Arg Ile-OtBu 892 Fmoc-Trp AspArg Leu-OtBu 893 Fmoc-Trp Arg Glu Leu-OtBu 894 Fmoc-Trp Glu Arg Leu-OtBu895 Fmoc-Phe Asp Arg Ile-OtBu 896 Fmoc-Phe Arg Glu Ile-OtBu 897 Fmoc-PheGlu Arg Ile-OtBu 898 Fmoc-Phe Asp Arg Leu-OtBu 899 Fmoc-Phe Arg GluLeu-OtBu 900 Fmoc-Phe Glu Arg Leu-OtBu 901 Fmoc-Trp Arg Asp Phe-OtBu 902Fmoc-Trp Arg Glu Phe-OtBu 903 Fmoc-Trp Glu Arg Phe-OtBu 904 Fmoc-Trp AspArg Tyr-OtBu 905 Fmoc-Trp Arg Glu Tyr-OtBu 906 Fmoc-Trp Glu Arg Tyr-OtBu907 Fmoc-Trp Arg Asp Thr(tBu)-OtBu  908 Fmoc-Trp Asp Arg Thr(tBu)-OtBu 909 Fmoc-Trp Arg Glu Thr(tBu)-OtBu  910 Fmoc-Trp Glu Arg Thr(tBu)-OtBu 911 Fmoc-Phe Arg Asp norLeu-OtBu 912 Fmoc-Phe Arg Glu norLeu-OtBu 913Boc-Phe Lys Asp Leu-OtB u 914 Boc-Phe Asp Lys Leu-OtBu 915 Boc-Phe LysGlu Leu-OtBu 916 Boc-Phe Glu Lys Leu-OtBu 917 Boc-Phe Lys Asp Ile-OtBu918 Boc-Phe Asp Lys Ile-OtBu 919 Boc-Phe Lys Glu Ile-OtBu 920 Boc-PheGlu Lys Ile-OtBu 921 Boc-Phe Lys Asp norLeu-OtBu 922 Boc-Phe Asp LysnorLeu-OtBu 923 Boc-Phe Lys Glu norLeu-OtBu 924 Boc-Phe Glu LysnorLeu-OtBu 925 Boc-Phe His Asp Leu-OtBu 926 Boc-Phe Asp His Leu-OtBu927 Boc-Phe His Glu Leu-OtBu 928 Boc-Phe Glu His Leu-OtBu 929 Boc-PheHis Asp Ile-OtBu 930 Boc-Phe Asp His Ile-OtBu 931 Boc-Phe His GluIle-OtBu 932 Boc-Phe Glu His Ile-OtBu 933 Boc-Phe His Asp norLeu-OtBu934 Boc-Phe Asp His norLeu-OtBu 935 Boc-Phe His Glu norLeu-OtBu 936Boc-Phe Glu His norLeu-OtBu 937 Boc-Lys(eBoc) Lys Asp Ser(tBu)-OtBu 938Boc-Lys(EBoc) Asp Lys Ser(tBu)-OtBu 939 Boc-Lys(eBoc) Lys GluSer(tBu)-OtBu 940 Boc-Lys(eBoc) Glu Lys Ser(tBu)-OtBu 941 Boc-Lys(eBoc)His Asp Ser(tBu)-OtBu 942 Boc-Lys(eBoc) Asp His Ser(tBu)-OtBu 943Boc-Lys(eBoc) His Glu Ser(tBu)-OtBu 944 Boc-Lys(EBoc) Glu HisSer(tBu)-OtBu 945

While the peptides of Table 8 are illustrated with particular protectinggroups, it is noted that these groups may be substituted with otherprotecting groups as described herein and/or one or more of the shownprotecting group can be eliminated.

4) Small Peptides Having Either an Acidic or Basic Amino Acid in theCenter Together with a Central Aliphatic Amino Acid.

In certain embodiments, the peptides of this invention range from fouramino acids to about ten amino acids. The terminal amino acids aretypically hydrophobic either because of a hydrophobic side chain orbecause the terminal amino acids bear one or more hydrophobic protectinggroups. End amino acids (X¹ and X⁴) are hydrophobic either because of ahydrophobic side chain or because the side chain or the C and/or Nterminus is blocked with one or more hydrophobic protecting group(s)(e.g., the N-terminus is blocked with Boc-, Fmoc-, Nicotinyl-, etc., andthe C-terminus blocked with (tBu)-OtBu, etc.). Typically, the centralportion of the peptide comprises a basic or acidic amino acid and analiphatic amino acid (e.g., in a 4 mer) or a basic domain or an acidicdomain and an aliphatic domain in a longer molecule.

These four-mers can be represented by Formula I in which X¹ and X⁴ arehydrophobic and/or bear hydrophobic protecting group(s) as describedherein and X² is acidic or basic while X³ is aliphatic or X² isaliphatic while X³ is acidic or basic. The peptide can be all L-aminoacids or include one, or more, or all D-amino acids.

Certain preferred peptides of this invention include, but are notlimited to the peptides shown in Table 9.

TABLE 9 Examples of certain preferred peptides havingeither an acidic or basic  amino acid in the center together with a central aliphatic amino acid. X¹ X² X³ X⁴ SEQ ID NOFmoc-Lys(εBoc) Leu Arg Ser(tBu)-OtBu 946 Fmoc-Lys(εBoc) Arg LeuSer(tBu)-OtBu 947 Fmoc-Lys(εBoc) Leu Arg Thr(tBu)-OtBu 948Fmoc-Lys(εBoc) Arg Leu Thr(tBu)-OtBu 949 Fmoc-Lys(εBoc) Glu LeuSer(tBu)-OtBu 950 Fmoc-Lys(εBoc) Leu Glu Ser(tBu)-OtBu 951Fmoc-Lys(εBoc) Glu Leu Thr(tBu)-OtBu 952 Fmoc-Lys(εBoc) Leu GluThr(tBu)-OtBu 953 Fmoc-Lys(εFmoc) Leu Arg Ser(tBu)-OtBu 954Fmoc-Lys(εFmoc) Leu Arg Thr(tBu)-OtBu 955 Fmoc-Lys(εFmoc) Glu LeuSer(tBu)-OtBu 956 Fmoc-Lys(εFmoc) Glu Leu Thr(tBu)-OtBu 957Boc-Lys(εFmoc) Glu Ile Thr(tBu)-OtBu 958 Boc-Lys(εFmoc) Leu ArgSer(tBu)-OtBu 959 Boc-Lys(εFmoc) Leu Arg Thr(tBu)-OtBu 960Boc-Lys(εFmoc) Glu Leu Ser(tBu)-OtBu 961 Boc-Lys(εFmoc) Glu LeuThr(tBu)-OtBu 962 Boc-Lys(εBoc) Leu Arg Ser(tBu)-OtBu 963 Boc-Lys(εBoc)Arg Phe Thr(tBu)-OtBu 964 Boc-Lys(εBoc) Leu Arg Thr(tBu)-OtBu 965Boc-Lys(εBoc) Glu Ile Thr(tBu) 966 Boc-Lys(εBoc) Glu Val Thr(tBu) 967Boc-Lys(εBoc) Glu Ala Thr(tBu) 968 Boc-Lys(εBoc) Glu Gly Thr(tBu) 969Boc-Lys(εBoc) Glu Leu Ser(tBu)-OtBu 970 Boc-Lys(εBoc) Glu LeuThr(tBu)-OtBu 971

While the peptides of Table 9 are illustrated with particular protectinggroups, it is noted that these groups may be substituted with otherprotecting groups as described herein and/or one or more of the shownprotecting group can be eliminated.

5) Small Peptides Having Either an Acidic or Basic Amino Acid in theCenter Together with a Central Aromatic Amino Acid.

In certain embodiments, the “small” peptides of this invention rangefrom four amino acids to about ten amino acids. The terminal amino acidsare typically hydrophobic either because of a hydrophobic side chain orbecause the terminal amino acids bear one or more hydrophobic protectinggroups end amino acids (X¹ and X⁴) are hydrophobic either because of ahydrophobic side chain or because the side chain or the C and/or Nterminus is blocked with one or more hydrophobic protecting group(s)(e.g., the N-terminus is blocked with Boc-, Fmoc-, Nicotinyl-, etc., andthe C-terminus blocked with (tBu)-OtBu, etc.). Typically, the centralportion of the peptide comprises a basic or acidic amino acid and anaromatic amino acid (e.g., in a 4 mer) or a basic domain or an acidicdomain and an aromatic domain in a longer molecule.

These four-mers can be represented by Formula I in which X¹ and X⁴ arehydrophobic and/or bear hydrophobic protecting group(s) as describedherein and X² is acidic or basic while X³ is aromatic or X² is aromaticwhile X³ is acidic or basic. The peptide can be all L-amino acids orinclude one, or more, or all D-amino acids. Five-mers can be representedby a minor modification of Formula I in which X⁵ is inserted as shown inTable 10 and in which X⁵ is typically an aromatic amino acid.

Certain preferred peptides of this invention include, but are notlimited to the peptides shown in Table 10.

TABLE 10 Examples of certain preferred peptides havingeither an acidic or basic amino acid in the centertogether with a central aromatic amino acid. SEQ ID X¹ X² X³ X⁵ X⁴ NOFmoc-Lys(εBoc) Arg Trp Tyr(tBu)-OtBu 972 Fmoc-Lys(εBoc) Trp ArgTyr(tBu)-OtBu 973 Fmoc-Lys(εBoc) Arg Tyr Trp-OtBu 974 Fmoc-Lys(εBoc) TyrArg Trp-OtBu 975 Fmoc-Lys(εBoc) Arg Tyr Trp Thr(tBu)-OtBu 976Fmoc-Lys(εBoc) Arg Tyr Thr(tBu)-OtBu 977 Fmoc-Lys(εBoc) Arg TrpThr(tBu)-OtBu 978 Fmoc-Lys(εFmoc) Arg Trp Tyr(tBu)-OtBu 979Fmoc-Lys(εFmoc) Arg Tyr Trp-OtBu 980 Fmoc-Lys(εFmoc) Arg Tyr Trp Thr(tBu)-OtBu 981 Fmoc-Lys(εFmoc) Arg Tyr Thr(tBu)-OtBu 982Fmoc-Lys(εFmoc) Arg Trp Thr(tBu)-OtBu 983 Boc-Lys(εFmoc) Arg TrpTyr(tBu)-OtBu 984 Boc-Lys(εFmoc) Arg Tyr Trp-OtBu 985 Boc-Lys(εFmoc) ArgTyr Trp Thr(tBu)-OtBu 986 Boc-Lys(εFmoc) Arg Tyr Thr(tBu)-OtBu 987Boc-Lys(εFmoc) Arg Tip Thr(tBu)-OtBu 988 Boc-Glu Lys(εFmoc) ArgTyr(tBu)-OtBu 989 Boc-Lys(εBoc) Arg Trp Tyr(tBu)-OtBu 990 Boc-Lys(εBoc)Arg Tyr Trp-OtBu 991 Boc-Lys(εBoc) Arg Tyr Trp Thr(tBu)-OtBu 992Boc-Lys(εBoc) Arg Tyr Thr(tBu)-OtBu 993 Boc-Lys(εBoc) Arg PheThr(tBu)-OtBu 994 Boc-Lys(εBoc) Arg Trp Thr(tBu)-OtBu 995

While the peptides of Table 10 are illustrated with particularprotecting groups, it is noted that these groups may be substituted withother protecting groups as described herein and/or one or more of theshown protecting group can be eliminated.

6) Small Peptides Having Aromatic Amino Acids or Aromatic Amino AcidsSeparated by Histidine(s) at the Center.

In certain embodiments, the peptides of this invention are characterizedby n electrons that are exposed in the center of the molecule whichallow hydration of the particle and that allow the peptide particles totrap pro-inflammatory oxidized lipids such as fatty acid hydroperoxidesand phospholipids that contain an oxidation product of arachidonic acidat the sn-2 position.

In certain embodiments, these peptides consist of a minimum of 4 aminoacids and a maximum of about 10 amino acids, preferentially (but notnecessarily) with one or more of the amino acids being theD-sterioisomer of the amino acid, with the end amino acids beinghydrophobic either because of a hydrophobic side chain or because theterminal amino acid(s) bear one or more hydrophobic blocking group(s),(e.g., an N-terminus blocked with Boc-, Fmoc-, Nicotinyl-, and the like,and a C-terminus blocked with (tBu)-OtBu groups and the like). Insteadof having an acidic or basic amino acid in the center, these peptidesgenerally have an aromatic amino acid at the center or have aromaticamino acids separated by histidine in the center of the peptide.

Certain preferred peptides of this invention include, but are notlimited to the peptides shown in Table 11.

TABLE 11 Examples of peptides having aromatic amino acids in the center or aromatic amino acids or aromatic domains separated by one   or more histidines. SEQ IDX¹ X² X³ X⁴ X⁵ NO Boc-Lys(εBoc) Phe Trp Phe Ser(tBu)-OtBu 996Boc-Lys(εBoc) Phe Trp Phe Thr(tBu)-OtBu 997 Boc-Lys(εBoc) Phe Tyr PheSer(tBu)-OtBu 998 Boc-Lys(εBoc) Phe Tyr Phe Thr(tBu)-OtBu 999Boc-Lys(εBoc) Phe His Phe Ser(tBu)-OtBu 1000 Boc-Lys(εBoc) Phe His PheThr(tBu)-OtBu 1001 Boc-Lys(εBoc) Val Phe Phe-Tyr Ser(tBu)-OtBu 1002Nicotinyl-Lys Phe Trp Phe Ser(tBu)-OtBu 1003 (εBoc) Nicotinyl-Lys PheTrp Phe Thr(tBu)-OtBu 1004 (εBoc) Nicotinyl-Lys Phe Tyr PheSer(tBu)-OtBu 1005 (εBoc) Nicotinyl-Lys Phe Tyr Phe Thr(tBu)-OtBu 1006(εBoc) Nicotinyl-Lys Phe His Phe Ser(tBu)-OtBu 1007 (εBoc) Nicotinyl-LysPhe His Phe Thr(tBu)-OtBu 1008 (εBoc) Boc-Leu Phe Trp Phe Thr(tBu)-OtBu1009 Boc-Leu Phe Trp Phe Ser(tBu)-OtBu 1010

While the peptides of Table 11 are illustrated with particularprotecting groups, it is noted that these groups may be substituted withother protecting groups as described herein and/or one or more of theshown protecting group can be eliminated.

7) Summary of Tripeptides and Tetrapeptides.

For the sake of clarity, a number of tripeptides and tetrapeptides ofthis invention are generally summarized below in Table 12.

TABLE 12 General structure of certain peptides of this invention. X¹ X²X³ X⁴ hydrophobic side Acidic or — hydrophobic side chain or hydrophobicBasic chain or hydrophobic protecting group(s) hydrophobic side BasicAcidic hydrophobic side chain or hydrophobic chain or hydrophobicprotecting group(s) protecting group(s) hydrophobic side Acidic Basichydrophobic side chain or hydrophobic chain or hydrophobic protectinggroup(s) protecting group(s) hydrophobic side Acidic or Aliphatichydrophobic side chain or hydrophobic Basic chain or hydrophobicprotecting group(s) protecting group(s) hydrophobic side AliphaticAcidic or hydrophobic side chain or hydrophobic Basic chain orhydrophobic protecting group(s) protecting group(s) hydrophobic sideAcidic or Aromatic hydrophobic side chain or hydrophobic Basic chain orhydrophobic protecting group(s) protecting group(s) hydrophobic sideAromatic Acidic or hydrophobic side chain or hydrophobic Basic chain orhydrophobic protecting group(s) protecting group(s) hydrophobic sideAromatic His hydrophobic side chain or hydrophobic Aromatic chain orhydrophobic protecting group(s) protecting group(s)

Where longer peptides are desired, X² and X³ can represent domains(e.g., regions of two or more amino acids of the specified type) ratherthan individual amino acids. Table 12 is intended to be illustrative andnot limiting. Using the teaching provided herein, other suitablepeptides can readily be identified.

8) Paired Amino Acids and Dipeptides.

In certain embodiments, this invention pertains to the discovery thatcertain pairs of amino acids, administered in conjunction with eachother or linked to form a dipeptide have one or more of the propertiesdescribed herein. Thus, without being bound to a particular theory, itis believed that when the pairs of amino acids are administered inconjunction with each other, as described herein, they are capableparticipating in or inducing the formation of micelles in vivo.

Similar to the other small peptides described herein, it is believedthat the pairs of peptides will associate in vivo, and demonstratephysical properties including high solubility in ethyl acetate (e.g.,greater than about 4 mg/mL), solubility in aqueous buffer at pH 7.0.Upon contacting phospholipids such as1,2-Dimyristoyl-sn-glycero-3-phosphocholine (DMPC), in an aqueousenvironment, it is believed the pairs of amino acids induce orparticipate in the formation of particles with a diameter ofapproximately 7.5 nm (.+−.0.1 nm), and/or induce or participate in theformation of stacked bilayers with a bilayer dimension on the order of3.4 to 4.1 nm with spacing between the bilayers in the stack ofapproximately 2 nm, and/or also induce or participate in the formationof vesicular structures of approximately 38 nm).

Moreover, it is further believed that the pairs of amino acids candisplay one or more of the following physiologically relevantproperties:

-   -   1. They convert pro-inflammatory HDL to anti-inflammatory HDL or        make anti-inflammatory HDL more anti-inflammatory;    -   2. They decrease LDL-induced monocyte chemotactic activity        generated by artery wall cells;    -   3. They stimulate the formation and cycling of pre-β HDL;    -   4. They raise HDL cholesterol; and/or    -   5. They increase HDL paraoxonase activity.

The pairs of amino acids can be administered as separate amino acids(administered sequentially or simultaneously, e.g. in a combinedformulation) or they can be covalently coupled directly or through alinker (e.g. a PEG linker, a carbon linker, a branched linker, astraight chain linker, a heterocyclic linker, a linker formed ofderivatized lipid, etc.). In certain embodiments, the pairs of aminoacids are covalently linked through a peptide bond to form a dipeptide.In various embodiments while the dipeptides will typically comprise twoamino acids each bearing an attached protecting group, this inventionalso contemplates dipeptides wherein only one of the amino acids bearsone or more protecting groups.

The pairs of amino acids typically comprise amino acids where each aminoacid is attached to at least one protecting group (e.g., a hydrophobicprotecting group as described herein). The amino acids can be in the Dor the L form. In certain embodiments, where the amino acids comprisingthe pairs are not attached to each other, each amino acid bears twoprotecting groups (e.g., such as molecules 1 and 2 in Table 13).

TABLE 13 Illustrative amino acid pairs of this invention.Amino Acid Pair/dipeptide 1. Boc-Arg-OtBu* 2. Boc-Glu-OtBu* 3.Boc-Phe-Arg-OtBu** 4. Boc-Glu-Leu-OtBu** 5. Boc-Arg-Glu-OtBu*** *Thiswould typically be administered in conjunction with a second aminoacid.**In certain embodiments, these dipeptides would be administered inconjunction with each other. ***In certain embodiments, this peptidewould be administered either alone or in combination with one of theother peptides described herein.

Suitable pairs of amino acids can readily be identified by providing thepair of protected amino acids and/or a dipeptide and then screening thepair of amino acids/dipeptide for one or more of the physical and/orphysiological properties described above. In certain embodiments, thisinvention excludes pairs of amino acids and/or dipeptides comprisingaspartic acid and phenylalanine. In certain embodiments, this inventionexcludes pairs of amino acids and/or dipeptides in which one amino acidis (−)-N-[(trans-4-isopropylcyclohexane)carbonyl]-D-phenylalanine(nateglinide).

In certain embodiments, the amino acids comprising the pair areindependently selected from the group consisting of an acidic amino acid(e.g., aspartic acid, glutamic acid, etc.), a basic amino acid (e.g.,lysine, arginine, histidine, etc.), and a non-polar amino acid (e.g.,alanine, valine, leucine, isoleucine, proline, phenylalanine,tryptophan, methionine, etc.). In certain embodiments, where the firstamino acid is acidic or basic, the second amino acid is non-polar andwhere the second amino acid is acidic or basic, the first amino acid isnon-polar. In certain embodiments, where the first amino acid is acidic,the second amino acid is basic, and vice versa. (see, e.g., Table 14).

Similar combinations can be obtained by administering pairs ofdipeptides. Thus, for example in certain embodiments, molecules 3 and 4in Table 13 would be administered in conjunction with each other.

TABLE 14 Certain generalized amino acid pairs/dipeptides. First Aminoacid Second Amino acid 1. Acidic Basic 2. Basic Acidic 3. AcidicNon-polar 4. Non-polar Acidic 5. Basic Non-polar 6. Non-polar Basic

It is noted that these amino acid pairs/dipeptides are intended to beillustrative and not limiting. Using the teaching provided herein othersuitable amino acid pairs/dipeptides can readily be determined.

E) Apo-J (G* Peptides).

It was a discovery of this invention that peptides that mimicking theamphipathic helical domains of apo J are capable of mitigating one ormore symptoms of atherosclerosis and/or other pathologies describedherein. Apolipoprotein J possesses a wide nonpolar face termed globularprotein-like, or G* amphipathic helical domains. The class G amphipathichelix is found in globular proteins, and thus, the name class G. Thisclass of amphipathic helix is characterized by a random distribution ofpositively charged and negatively charged residues on the polar facewith a narrow nonpolar face. Because of the narrow nonpolar face thisclass does not readily associate with phospholipids. The G* ofamphipathic helix possesses similar, but not identical, characteristicsto the G amphipathic helix. Similar to the class G amphipathic helix,the G* class peptides possesses a random distribution of positively andnegatively charged residues on the polar face. However, in contrast tothe class G amphipathic helix which has a narrow nonpolar face, thisclass has a wide nonpolar face that allows this class to readily bindphospholipid and the class is termed G* to differentiate it from the Gclass of amphipathic helix.

A number of suitable G* amphipathic peptides are described in copendingapplications U.S. Ser. No. 10/120,508, filed Apr. 5, 2002, U.S. Ser. No.10/520,207, filed Apr. 1, 2003, and PCT Application PCT/US03/09988,filed Apr. 1, 2003. In addition, a variety of suitable peptides of thisinvention that are related to G* amphipathic helical domains of apo Jare illustrated in Table 15.

TABLE 15 Certain illustrative peptides for use inthis invention related to G* amphipathic helical domains of apo J.Amino Acid Sequence SEQ ID NO LLEQLNEQFNWVSRLANLTQGE 1011LLEQLNEQFNWVSRLANL 1012 NELQEMSNQGSKYVNKEIQNAVNGV 1013IQNAVNGVKQIKTLIEKTNEE 1014 RKTLLSNLEEAKKKKEDALNETRESETKLKEL 1015PGVCNETMMALWEECK 1016 PCLKQTCMKFYARVCR 1017 ECKPCLKQTCMKFYARVCR 1018LVGRQLEEFL 1019 MNGDRIDSLLEN 1020 QQTHMLDVMQD 1021 FSRASSIIDELFQD 1022PFLEMIHEAQQAMDI 1023 PTEFIREGDDD 1024 RMKDQCDKCREILSV 1025PSQAKLRRELDESLQVAERLTRKYNELLKSYQ 1026 LLEQLNEQFNWVSRLANLTEGE 1027DQYYLRVTTVA 1028 PSGVTEVVVKLFDS 1029 PKFMETVAEKALQEYRKKHRE 1030

The peptides of this invention, however, are not limited to G* variantsof apo J. Generally speaking G* domains from essentially any otherprotein preferably apo proteins are also suitable. The particularsuitability of such proteins can readily be determined using assays forprotective activity (e.g., protecting LDL from oxidation, and the like),e.g. as illustrated herein in the Examples. Some particularly preferredproteins include G* amphipathic helical domains or variants thereof(e.g., conservative substitutions, and the like) of proteins including,but not limited to apo AI, apo AIV, apo E, apo CII, apo CIII, and thelike.

Certain preferred peptides for related to G* amphipathic helical domainsrelated to apoproteins other than apo J are illustrated in Table 16.

TABLE 16 Peptides for use in this invention related toG* amphipathic helical domains related to apoproteins other than apo J.SEQ ID Amino Acid Sequence  NO WDRVKDLATVYVDVLKDSGRDYVSQF 1031(Related to the 8 to 33 region of apo AI) VATVMWDYFSQLSNNAKEAVEHLQK 1032(Related to the 7 to 31 region of apo AIV) RWELALGRFWDYLRWVQTLSEQVQEEL1033 (Related to the 25 to 51 region of apo E)LSSQVTQELRALMDETMKELKELKAYKSELEEQLT 1034(Related to the 52 to 83 region of apo E) ARLSKELQAAQARLGADMEDVCGRLV 1035 (Related to the 91 to 116 region of apo E)VRLASHLRKLRKRLLRDADDLQKRLA  1036(Related to the135 to 160 region of apo E) PLVEDMQRQWAGLVEKVQA 1037(267 to 285 of apo E.27) MSTYTGIFTDQVLSVLK 1038(Related to the 60 to 76 region of apo CII) LLSFMQGYMKHATKTAKDALSS 1039(Related to the 8 to 29 region of apo CIII)

Additional illustrative G* peptides are shown in Table 17.

TABLE 17 Additional illustrative G* peptides. SEQ ID Peptide NOAc-Lys-Trp-Ile-Tyr-His-Leu-Thr-Glu-Gly- 1040Ser-Thr- Asp-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Trp-Phe-Tyr-His-Leu-Thr-Glu-Gly- 1041Ser-Thr- Asp-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Trp-Leu-Tyr-His-Leu-Thr-Glu-Gly- 1042Ser-Thr- Asp-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Trp-Val-Tyr-His-Leu-Thr-Glu-Gly- 1043Ser-Thr- Asp-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Tyr-Ile-Trp-His-Leu-Thr-Glu-Gly- 1044Ser-Thr- Asp-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Trp-Ile-Tyr-His-Phe-Thr-Glu-Gly- 1045Ser-Thr- Asp-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Trp-Phe-Tyr-His-Ile-Thr-Glu-Gly- 1046Ser-Thr- Asp-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Trp-Leu-Tyr-His-Val-Thr-Glu-Gly- 1047Ser-Thr- Asp-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Trp-Val-Tyr-His-Tyr-Thr-Glu-Gly- 1048Ser-Thr- Asp-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Tyr-Ile-Trp-His-Phe-Thr-Glu-Gly- 1049Ser-Thr- Asp-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Tyr-Ile-Trp-His-Ile-Thr-Glu-Gly- 1050Ser-Thr- Asp-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Tyr-Ile-Trp-His-Val-Thr-Glu-Gly- 1051Ser-Thr- Asp-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Tyr-Ile-Trp-His-Tyr-Thr-Glu-Gly- 1052Ser-Thr- Asp-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Phe-Ile-Trp-His-Leu-Thr-Glu-Gly- 1053Ser-Thr- Asp-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Leu-Ile-Trp-His-Leu-Thr-Glu-Gly- 1054Ser-Thr- Asp-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Ile-Ile-Trp-His-Leu-Thr-Glu-Gly- 1055Ser-Thr- Asp-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Tyr-Ile-Trp-Phe-Leu-Thr-Glu-Gly- 1056Ser-Thr- Asp-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Trp-Ile-Tyr-Phe-Leu-Thr-Glu-Gly- 1057Ser-Thr- Asp-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Trp-Ile-Tyr-Leu-Leu-Thr-Glu-Gly- 1058Ser-Thr- Asp-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Trp-Ile-Tyr-His-Phe-Thr-Glu-Gly- 1059Ser-Thr- Asp-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Trp-Ile-Tyr-His-Tyr-Thr-Glu-Gly- 1060ASer-Thr- Asp-Leu-rg-Thr-Glu-Gly-NH₂Ac-Lys-Trp-Ile-Tyr-His-Ile-Thr-Glu-Gly- 1061Ser-Thr- Asp-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Trp-Ile-Tyr-His-Leu-Ser-Glu-Gly- 1062Ser-Thr- Asp-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Trp-Ile-Tyr-His-Leu-Thr-Asp-Gly- 1063Ser-Thr- Asp-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Trp-Ile-Tyr-His-Leu-Thr-Glu-Gly- 1064Thr- Ser-Asp-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Trp-Ile-Tyr-His-Leu-Thr-Glu-Gly- 1065Ser- Thr-Glu-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Trp-Ile-Tyr-His-Leu-Thr-Glu-Gly- 1066Ser-Thr- Asp-Phe-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Trp-Ile-Tyr-His-Leu-Thr-Glu-Gly- 1067Ser-Thr- Asp-Tyr-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Trp-Ile-Tyr-His-Leu-Thr-Glu-Gly- 1068Ser-Thr- Asp-Ile-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Trp-Ile-Tyr-His-Leu-Thr-Glu-Gly- 1069Ser-Thr- Asp-Val-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Trp-Ile-Tyr-His-Leu-Thr-Glu-Gly- 1070Ser-Thr- Asp-Leu-Lys-Thr-Glu-Gly-NH₂Ac-Lys-Trp-Ile-Tyr-His-Leu-Thr-Glu-Gly- 1071Ser-Thr- Asp-Leu-Arg-Ser-Glu-Gly-NH₂Ac-Lys-Trp-Ile-Tyr-His-Leu-Thr-Glu-Gly- 1072Ser-Thr- Asp-Leu-Arg-Thr-Asp-Gly-NH₂Ac-Lys-Trp-Ile-Tyr-His-Leu-Thr-Glu-Gly- 1073Ser-Thr- Asp-Ile-Lys-Thr-Glu-Gly-NH₂Ac-Lys-Trp-Ile-Tyr-His-Leu-Thr-Glu-Gly- 1074Ser-Thr- Asp-Ile-Arg-Ser-Glu-Gly-NH₂Ac-Lys-Trp-Ile-Tyr-His-Leu-Thr-Glu-Gly- 1075Ser-Thr- Asp-Ile-Lys-Ser-Glu-Gly-NH₂Ac-Lys-Trp-Ile-Tyr-His-Leu-Thr-Glu-Gly- 1076Ser-Thr- Asp-Ile-Lys-Ser-Asp-Gly-NH₂Ac-Arg-Trp-Ile-Tyr-His-Leu-Thr-Glu-Gly- 1077Ser-Thr- Asp-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Arg-Tyr-Ile-Trp-His-Leu-Thr-Glu-Gly- 1078Ser-Thr- Asp-Ile-Arg-Thr-Glu-Gly-NH₂Ac-Arg-Trp-Ile-Tyr-His-Leu-Thr-Glu-Gly- 1079Ser-Thr- Asp-Ile-Arg-Thr-Asp-Gly-NH₂Ac-Arg-Trp-Ile-Phe-His-Leu-Thr-Glu-Gly- 1080Ser-Thr- Asp-Ile-Arg-Thr-Glu-Gly-NH₂Ac-Arg-Trp-Ile-Tyr-His-Leu-Thr-Glu-Gly- 1081Ser-Thr- Asp-Leu-Lys-Thr-Glu-Gly-NH₂Ac-Arg-Trp-Ile-Tyr-His-Leu-Thr-Asp-Gly- 1082Ser-Thr- Asp-Ile-Arg-Thr-Glu-Gly-NH₂Ac-Arg-Trp-Ile-Tyr-His-Leu-Thr-Asp-Gly- 1083Ser-Thr- Asp-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Arg-Trp-Ile-Tyr-Phe-Leu-Thr-Glu-Gly- 1084Ser-Thr- Asp-Ile-Arg-Thr-Glu-Gly-NH₂Ac-Arg-Trp-Ile-Tyr-Phe-Leu-Thr-Glu-Gly- 1085Ser-Thr- Asp-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Trp-Phe-Tyr-His-Leu-Thr-Glu-Gly- 1086Ser-Thr- Asp-Phe-Arg-Thr-Glu-Gly-NH₂Ac-Arg-Trp-Phe-Tyr-His-Leu-Thr-Glu-Gly- 1087Ser-Thr- Asp-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Trp-Ile-Phe-His-Leu-Thr-Glu-Gly- 1088Ser-Thr- Asp-Ile-Arg-Thr-Asp-Gly-NH₂Ac-Arg-Trp-Ile-Tyr-His-Leu-Thr-Glu-Gly- 1089Ser-Thr- Asp-Ile-Arg-Thr-Asp-Gly-NH₂Ac-Arg-Trp-Ile-Tyr-His-Leu-Thr-Glu-Gly- 1090Ser-Thr- Asp-Leu-Arg-Thr-Asp-Gly-NH₂Ac-Lys-Trp-Ile-Tyr-His-Leu-Thr-Glu-Gly- 1091Ser-Thr- Asp-Ile-Lys-Thr-Glu-Gly-NH₂Ac-Lys-Trp-Ile-Tyr-His-Leu-Thr-Glu-Gly- 1092Ser-Thr- Asp-Ile-Lys-Thr-Asp-Gly-NH₂Ac-Lys-Trp-Ile-Tyr-His-Leu-Thr-Glu-Gly- 1093Ser-Thr- Asp-Phe-Lys-Thr-Glu-Gly-NH₂Ac-Lys-Trp-Ile-Tyr-His-Leu-Thr-Glu-Gly- 1094Ser-Thr- Asp-Tyr-Lys-Thr-Glu-Gly-NH₂Ac-Lys-Trp-Ile-Tyr-His-Leu-Thr-Glu-Gly- 1095Ser-Thr- Asp-Ile-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Trp-Phe-Tyr-His-Phe-Thr-Glu-Gly- 1096Ser-Thr- Asp-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Arg-Trp-Phe-Tyr-His-Phe-Thr-Glu-Gly- 1097Ser-Thr- Asp-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Trp-Phe-Tyr-His-Phe-Thr-Glu-Gly- 1098Ser-Thr- Asp-Phe-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Trp-Phe-Tyr-His-Phe-Thr-Asp-Gly- 1099Ser-Thr- Asp-Ile-Arg-Thr-Glu-Gly-NH₂Ac-Arg-Trp-Phe-Tyr-His-Phe-Thr-Glu-Gly- 1100Ser-Thr- Asp-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Arg-Trp-Phe-Tyr-His-Phe-Thr-Glu-Gly- 1101Ser-Thr- Asp-Phe-Arg-Thr-Glu-Gly-NH₂Ac-Arg-Trp-Phe-Tyr-His-Phe-Thr-Glu-Gly- 1102Ser-Thr- Asp-Phe-Arg-Thr-Asp-Gly-NH₂Ac-Glu-Lys-Cys-Val-Glu-Glu-Phe-Lys-Ser- 1103Leu-Thr- Ser-Cys-Leu-Asp-Ser-Lys-Ala- Phe-NH₂Ac-Asp-Lys-Cys-Val-Glu-Glu-Phe-Lys-Ser- 1104Leu-Thr- Ser-Cys-Leu-Asp-Ser-Lys-Ala- Phe-NH₂Ac-Glu-Lys-Cys-Val-Asp-Glu-Phe-Lys-Ser- 1105Leu-Thr- Ser-Cys-Leu-Asp-Ser-Lys-Ala- Phe-NH₂Ac-Glu-Lys-Cys-Val-Glu-Asp-Phe-Lys-Ser- 1106Leu-Thr- Ser-Cys-Leu-Asp-Ser-Lys-Ala- Phe-NH₂Ac-Glu-Arg-Cys-Val-Glu-Glu-Phe-Lys-Ser- 1107Leu-Thr- Ser-Cys-Leu-Asp-Ser-Lys-Ala- Phe-NH₂Ac-Asp-Lys-Cys-Val-Asp-Asp-Phe-Lys-Ser- 1108Leu-Thr- Ser-Cys-Leu-Asp-Ser-Lys-Ala- Phe-NH₂Ac-Asp-Arg-Cys-Val-Glu-Glu-Phe-Lys-Ser- 1109Leu-Thr- Ser-Cys-Leu-Asp-Ser-Lys-Ala- Phe-NH₂Ac-Glu-Arg-Cys-Val-Asp-Asp-Phe-Lys-Ser- 1110Leu-Thr- Ser-Cys-Leu-Asp-Ser-Lys-Ala- Phe-NH₂Ac-Glu-Lys-Cys-Val-Glu-Glu-Phe-Lys-Ser- 1111Phe-Thr- Ser-Cys-Leu-Asp-Ser-Lys-Ala- Phe-NH₂Ac-Glu-Lys-Cys-Val-Glu-Glu-Phe-Lys-Ser- 1112Ile-Thr- Ser-Cys-Leu-Asp-Ser-Lys-Ala- Phe-NH₂Ac-Glu-Lys-Cys-Val-Glu-Glu-Phe-Lys-Ser- 1113Val-Thr- Ser-Cys-Leu-Asp-Ser-Lys-Ala- Phe-NH₂Ac-Glu-Arg-Cys-Val-Glu-Glu-Phe-Lys-Ser- 1114Tyr-Thr- Ser-Cys-Leu-Asp-Ser-Lys-Ala- Phe-NH₂Ac-Glu-Arg-Cys-Val-Glu-Glu-Phe-Lys-Ser- 1115Phe-Thr- Ser-Cys-Leu-Asp-Ser-Lys-Ala- Phe-NH₂Ac-Glu-Arg-Cys-Val-Glu-Glu-Phe-Lys-Ser- 1116Ile-Thr- Ser-Cys-Leu-Asp-Ser-Lys-Ala- Phe-NH₂Ac-Glu-Arg-Cys-Val-Glu-Glu-Phe-Lys-Ser- 1117Val-Thr- Ser-Cys-Leu-Asp-Ser-Lys-Ala- Phe-NH₂Ac-Glu-Arg-Cys-Val-Glu-Glu-Phe-Lys-Ser- 1118Tyr-Thr- Ser-Cys-Leu-Asp-Ser-Lys-Ala- Phe-NH₂Ac-Glu-Lys-Cys-Val-Glu-Glu-Phe-Lys-Ser- 1119Phe-Thr -Thr-Cys-Leu-Asp-Ser-Lys-Ala- Phe-NH₂Ac-Glu-Lys-Cys-Val-Glu-Glu-Phe-Lys-Ser- 1120Ile-Ser- Ser-Cys-Leu-Asp-Ser-Lys-Ala- Phe-NH₂Ac-Glu-Lys-Cys-Val-Glu-Glu-Phe-Lys-Ser- 1121Val-Ser- Thr-Cys-Leu-Asp-Ser-Lys-Ala- Phe-NH₂Ac-Glu-Lys-Cys-Val-Glu-Glu-Phe-Lys-Ser- 1122Tyr-Thr- Ser-Cys-Leu-Asp-Ser-Lys-Ala- Phe-NH₂Ac-Glu-Lys-Cys-Val-Glu-Glu-Phe-Lys-Ser- 1123Phe-Thr-Thr-Cys-Leu-Asp-Ser-Lys-Ala- Phe-NH₂Ac-Glu-Lys-Cys-Val-Glu-Glu-Phe-Lys-Ser- 1124Phe-Ser-Ser-Cys-Leu-Asp-Ser-Lys-Ala- Phe-NH₂Ac-Glu-Lys-Cys-Val-Glu-Glu-Phe-Lys-Ser- 1125Phe-Thr- Ser-Cys-Leu-Asp-Ser-Lys-Ala- Phe-NH₂Ac-Glu-Lys-Cys-Val-Glu-Glu-Phe-Lys-Ser- 1126Phe-Thr- Ser-Cys-Leu-Asp-Ser-Lys-Ala- Phe-NH₂Ac-Glu-Lys-Cys-Val-Glu-Glu-Phe-Lys-Ser- 1127Phe-Thr- Ser-Cys-Leu-Asp-Ser-Lys-Ala- Phe-NH₂Ac-Glu-Lys-Cys-Val-Glu-Glu-Phe-Lys-Ser- 1128Phe-Thr- Ser-Cys-Phe-Asp-Ser-Lys-Ala- Phe-NH₂Ac-Glu-Lys-Cys-Val-Glu-Glu-Phe-Lys-Ser- 1129Phe-Thr- Ser-Cys-Phe-Glu-Ser-Lys-Ala- Phe-NH₂Ac-Glu-Lys-Cys-Val-Glu-Glu-Phe-Lys-Ser- 1130Phe-Thr- Ser-Cys-Leu-Glu-Ser-Lys-Ala- Phe-NH₂Ac-Glu-Lys-Cys-Val-Glu-Glu-Phe-Lys-Ser- 1131Phe-Thr- Ser-Cys-Ile-Asp-Ser-Lys-Ala- Phe-NH₂Ac-Glu-Lys-Cys-Val-Glu-Glu-Leu-Lys-Ser- 1132Phe-Thr- Ser-Cys-Phe-Asp-Ser-Lys-Ala- Phe-NH₂Ac-Asp-Lys-Cys-Val-Glu-Glu-Phe-Lys-Ser- 1133Phe-Thr- Ser-Cys-Phe-Asp-Ser-Lys-Ala- Phe-NH₂Ac-Asp-Lys-Cys-Val-Glu-Glu-Phe-Lys-Ser- 1134Phe-Thr- Ser-Cys-Phe-Glu-Ser-Lys-Ala- Phe-NH₂Ac-Glu-Arg-Cys-Val-Glu-Glu-Phe-Lys-Ser- 1135Phe-Thr- Ser-Cys-Phe-Asp-Ser-Lys-Ala- Phe-NH₂Ac-Glu-Lys-Cys-Phe-Glu-Glu-Phe-Lys-Ser- 1136Phe-Thr- Ser-Cys-Phe-Asp-Ser-Lys-Ala- Phe-NH₂Ac-Glu-Lys-Cys-Phe-Glu-Glu-Phe-Lys-Ser- 1137Phe-Thr- Ser-Cys-Phe-Glu-Ser-Lys-Ala- Phe-NH₂Ac-Glu-Lys-Cys-Val-Glu-Glu-Phe-Lys-Ser- 1138Phe-Ser- Ser-Cys-Phe-Glu-Ser-Lys-Ala- Phe-NH₂Ac-Glu-Lys-Cys-Val-Glu-Glu-Phe-Lys-Ser- 1139Phe-Gln- Ser-Cys-Phe-Asp-Ser-Lys-Ala- Phe-NH₂Ac-Glu-Lys-Cys-Phe-Glu-Glu-Phe-Lys-Ser- 1140Phe-Gln- Ser-Cys-Phe-Asp-Ser-Lys-Ala- Phe-NH₂Ac-Glu-Lys-Cys-Val-Glu-Glu-Phe-Lys-Gln- 1141Phe-Thr- Ser-Cys-Phe-Asp-Ser-Lys-Ala- Phe-NH₂Ac-Glu-Lys-Cys-Val-Glu-Glu-Phe-Lys-Gln- 1142Leu-Thr- Ser-Cys-Leu-Asp-Ser-Lys-Ala- Phe-NH₂Ac-Glu-Lys-Cys-Phe-Glu-Glu-Phe-Lys-Ser- 1143Phe-Gln- Ser-Cys-Leu-Asp-Ser-Lys-Ala- Phe-NH₂Ac-Glu-Lys-Cys-Val-Glu-Glu-Phe-Lys-Gln- 1144Phe-Thr- Ser-Cys-Phe-Asp-Ser-Lys-Ala- Phe-NH₂Ac-Glu-Lys-Cys-Val-Glu-Glu-Phe-Lys-Ser- 1145Phe-Thr- Ser-Cys-Phe-Glu-Ser-Lys-Ala- Phe-NH₂Ac-Glu-Arg-Cys-Phe-Glu-Glu-Phe-Lys-Ser- 1146Phe-Thr- Ser-Cys-Phe-Asp-Ser-Lys-Ala- Phe-NH₂Ac-Asp-Lys-Cys-Phe-Glu-Glu-Phe-Lys-Ser- 1147Phe-Thr- Ser-Cys-Phe-Asp-Ser-Lys-Ala- Phe-NH₂Ac-Glu-Arg-Cys-Val-Glu-Glu-Phe-Lys-Ser- 1148Leu-Thr- Ser-Cys-Leu-Glu-Ser-Lys-Ala- Phe-NH₂Ac-Glu-Lys-Cys-Val-Glu-Glu-Phe-Lys-Ser- 1149Leu-Thr- Ser-Cys-Leu-Asp-Ser-Lys-Phe- Phe-NH₂Ac-Glu-Lys-Cys-Phe-Glu-Glu-Phe-Lys-Ser- 1150Phe-Thr- Ser-Cys-Phe-Asp-Ser-Lys-Phe- Phe-NH₂Ac-Asp-Lys-Cys-Phe-Glu-Glu-Phe-Lys-Ser- 1151Phe-Thr- Ser-Cys-Leu-Asp-Ser-Lys-Phe- Phe-NH₂Ac-Asp-Lys-Cys-Phe-Glu-Glu-Phe-Lys-Ser- 1152Phe-Thr- Ser-Cys-Leu-Glu-Ser-Lys-Phe- Phe-NH₂Ac-Asp-Lys-Cys-Phe-Glu-Glu-Leu-Lys-Ser- 1153Phe-Thr- Ser-Cys-Leu-Asp-Ser-Lys-Phe- Phe-NH₂Ac-Glu-Arg-Cys-Phe-Glu-Glu-Phe-Lys-Ser- 1154Phe-Thr- Ser-Cys-Leu-Asp-Ser-Lys-Phe- Phe-NH₂Ac-Glu-Lys-Ala-Val-Glu-Glu-Phe-Lys-Ser- 1155Phe-Thr- Ser-Cys-Leu-Asp-Ser-Lys-Ala- Phe-NH₂Ac-Asp-Lys-Ala-Val-Glu-Glu-Phe-Lys-Ser- 1156Phe-Thr- Ser-Cys-Leu-Asp-Ser-Lys-Phe- Phe-NH₂Ac-Glu-Lys-Ala-Val-Glu-Glu-Phe-Lys-Ser- 1157Phe-Thr- Ser-Ala-Leu-Asp-Ser-Lys-Ala- Phe-NH₂Ac-Asp-Lys-Ala-Val-Glu-Glu-Phe-Lys-Ser- 1158Phe-Thr- Ser-Ala-Leu-Asp-Ser-Lys-Ala- Phe-NH₂Ac-Asp-Arg-Ala-Phe-Glu-Glu-Phe-Lys-Ser- 1159Phe-Thr- Ser-Cys-Leu-Asp-Ser-Lys-Phe- Phe-NH₂Ac-Asp-Arg-Ala-Phe-Glu-Glu-Phe-Lys-Ser- 1160Phe-Thr- Ser-Ala-Leu-Asp-Ser-Lys-Phe- Phe-NH₂Ac-Asp-Lys-Cys-Phe-Glu-Glu-Phe-Lys-Ser- 1161Phe-Thr- Ser-Cys-Phe-Glu-Ser-Lys-Phe- Phe-NH₂Ac-Glu-Lys-Cys-Tyr-Glu-Glu-Phe-Lys-Ser- 1162Phe-Thr- Ser-Cys-Leu-Asp-Ser-Lys-Phe- Phe-NH₂Ac-Asp-Lys-Cys-Trp-Glu-Glu-Phe-Lys-Ser- 1163Phe-Thr- Ser-Cys-Leu-Asp-Ser-Lys-Phe- Phe-NH₂Ac-Glu-Lys-Cys-Phe-Glu-Glu-Phe-Lys-Ser- 1164Tyr-Thr- Ser-Cys-Leu-Asp-Ser-Lys-Phe- Phe-NH₂Ac-Glu-Lys-Cys-Phe-Glu-Glu-Phe-Lys-Ser- 1165Trp-Thr- Ser-Cys-Leu-Asp-Ser-Lys-Phe- Phe-NH₂Ac-Glu-Lys-Cys-Val-Glu-Glu-Phe-Lys-Ser- 1166Trp-Thr- Ser-Cys-Leu-Asp-Ser-Lys-Ala- Phe-NH₂Ac-Asp-Lys-Cys-Phe-Glu-Glu-Phe-Lys-Ser- 1167Trp-Thr- Ser-Cys-Leu-Asp-Ser-Lys-Ala- Phe-NH₂

Other suitable peptides include, but are not limited to the peptides ofTable 18.

TABLE 18 Illustrative peptides having an improved hydrophobic phase.SEQ ID Name Sequence NO V2W3A5F1017-D-4F Ac-Asp-Val-Trp-Lys-Ala-Ala-1168 Tyr-Asp-Lys-Phe-Ala-Glu-Lys- Phe-Lys-Glu-Phe-Phe-NH₂ V2W3F10-D-4FAc-Asp-Val-Trp-Lys-Ala-Phe- 1169 Tyr-Asp-Lys-Phe-Ala-Glu-Lys-Phe-Lys-Glu-Ala-Phe-NH₂ W3-D-4F Ac-Asp-Phe-Trp-Lys-Ala-Phe- 1170Tyr-Asp-Lys-Val-Ala-Glu-Lys- Phe-Lys-Glu-Ala-Phe-NH₂

The peptides described here (V2W3A5F10,17-D-4F; V2W3F10-D-4F; W3-D-4F)may be more potent than the original D-4F.

Still other suitable peptides include, but are not limited to:P¹-Dimethyltyrosine-D-Arg-Phe-Lys-P² (SEQ ID NO: 1171) andP¹-Dimethyltyrosine-Arg-Glu-Leu-P² (SEQ ID NO: 1206) where P1 and P2 areprotecting groups as described herein. In certain embodiments, thesepeptides include, but are not limited toBocDimethyltyrosine-D-Arg-Phe-Lys(OtBu) (SEQ ID NO: 1207) andBocDimethyltyrosine-Arg-Glu-Leu(OtBu) (SEQ ID NO: 1208).

In certain embodiments, the peptides of this invention include peptidescomprising or consisting of the amino acid sequence LAEYHAK (SEQ IDNO:1172) comprising at least one D amino acid and/or at least one or twoterminal protecting groups. In certain embodiments, this inventionincludes a peptide that ameliorates one or more symptoms of aninflammatory condition, wherein the peptide: ranges in length from about3 to about 10 amino acids; comprises an amino acid sequence where thesequence comprises acidic or basic amino acids alternating with aromaticor hydrophobic amino acids; comprises hydrophobic terminal amino acidsor terminal amino acids bearing a hydrophobic protecting group; is notthe sequence LAEYHAK (SEQ ID NO:1173) comprising all L amino acids;where the peptide converts pro-inflammatory HDL to anti-inflammatory HDLand/or makes anti-inflammatory HDL more anti-inflammatory.

It is also noted that the peptides listed in the Tables herein are notfully inclusive. Using the teaching provided herein, other suitablepeptides can routinely be produced (e.g. by conservative orsemi-conservative substitutions (e.g. D replaced by E), extensions,deletions, and the like). Thus, for example, one embodiment utilizestruncations of any one or more of peptides identified by SEQ IDNos:1011-1039.

Longer peptides are also suitable. Such longer peptides may entirelyform a class G or G* amphipathic helix, or the G amphipathic helix(helices) can form one or more domains of the peptide. In addition, thisinvention contemplates multimeric versions of the peptides. Thus, forexample, the peptides illustrated in the tables herein can be coupledtogether (directly or through a linker (e.g. a carbon linker, or one ormore amino acids) with one or more intervening amino acids). Suitablelinkers include, but are not limited to Proline (-Pro-), Gly₄Ser₃ (SEQID NO:1174), and the like. Thus, one illustrative multimeric peptideaccording to this invention is (D-J336)-P-(D-J336) (i.e.Ac-L-L-E-Q-L-N-E-Q-F-N-W-V-S-R-L-A-N-L-T-Q-G-E-P-L-L-E-Q-L-N-E-Q-F-N-W-V-S-R-L-A-N-L-T-Q-G-E-NH₂,SEQ ID NO:1175).

This invention also contemplates the use of “hybrid” peptides comprisinga one or more G or G* amphipathic helical domains and one or more classA amphipathic helices. Suitable class A amphipathic helical peptides aredescribed in PCT publication WO 02/15923. Thus, by way of illustration,one such “hybrid” peptide is (D-J336)-Pro-4F) (i.e.Ac-L-L-E-Q-L-N-E-Q-F-N-W-V-S-R-L-A-N-L-T-Q-G-E-P-D-W-F-K-A-F-Y-D-K-V-A-E-K-F-K-E-A-F-NH₂,SEQ ID NO: 1176), and the like.

Using the teaching provided herein, one of skill can routinely modifythe illustrated amphipathic helical peptides to produce other suitableapo J variants and/or amphipathic G and/or A helical peptides of thisinvention. For example, routine conservative or semi-conservativesubstitutions (e.g., E for D) can be made of the existing amino acids.The effect of various substitutions on lipid affinity of the resultingpeptide can be predicted using the computational method described byPalgunachari et al. (1996) Arteriosclerosis, Thrombosis, & VascularBiology 16: 328-338. The peptides can be lengthened or shortened as longas the class helix structure(s) are preserved. In addition,substitutions can be made to render the resulting peptide more similarto peptide(s) endogenously produced by the subject species.

While, in preferred embodiments, the peptides of this invention utilizenaturally-occurring amino acids or D forms of naturally occurring aminoacids, substitutions with non-naturally occurring amino acids (e.g.,methionine sulfoxide, methionine methylsulfonium, norleucine,episilon-aminocaproic acid, 4-aminobutanoic acid,tetrahydroisoquinoline-3-carboxylic acid, 8-aminocaprylic acid,4-aminobutyric acid, Lys(N(epsilon)-trifluoroacetyl), α-aminoisobutyricacid, and the like) are also contemplated.

New peptides can be designed and/or evaluated using computationalmethods. Computer programs to identify and classify amphipathic helicaldomains are well known to those of skill in the art and many have beendescribed by Jones et al. (1992) J. Lipid Res. 33: 287-296). Suchprograms include, but are not limited to the helical wheel program(WHEEL or WHEEL/SNORKEL), helical net program (HELNET, HELNET/SNORKEL,HELNET/Angle), program for addition of helical wheels (COMBO orCOMBO/SNORKEL), program for addition of helical nets (COMNET,COMNET/SNORKEL, COMBO/SELECT, COMBO/NET), consensus wheel program(CONSENSUS, CONSENSUS/SNORKEL), and the like.

F) Blocking Groups and D Residues.

While the various peptides and/or amino acid pairs described herein maybe shown with no protecting groups, in certain embodiments (e.g.particularly for oral administration), they can bear one, two, three,four, or more protecting groups. The protecting groups can be coupled tothe C- and/or N-terminus of the peptide(s) and/or to one or moreinternal residues comprising the peptide(s) (e.g., one or more R-groupson the constituent amino acids can be blocked). Thus, for example, incertain embodiments, any of the peptides described herein can bear, e.g.an acetyl group protecting the amino terminus and/or an amide groupprotecting the carboxyl terminus. One example of such a “dual protectedpeptide is Ac-L-L-E-Q-L-N-E-Q-F-N-W-V-S-R-L-A-N-L-T-Q-G-E-NH₂ (SEQ IDNO:1011 with blocking groups), either or both of these protecting groupscan be eliminated and/or substituted with another protecting group asdescribed herein.

Without being bound by a particular theory, it was a discovery of thisinvention that blockage, particularly of the amino and/or carboxyltermini of the subject peptides of this invention greatly improves oraldelivery and significantly increases serum half-life.

A wide number of protecting groups are suitable for this purpose. Suchgroups include, but are not limited to acetyl, amide, and alkyl groupswith acetyl and alkyl groups being particularly preferred for N-terminalprotection and amide groups being preferred for carboxyl terminalprotection. In certain particularly preferred embodiments, theprotecting groups include, but are not limited to alkyl chains as infatty acids, propeonyl, formyl, and others. Particularly preferredcarboxyl protecting groups include amides, esters, and ether-formingprotecting groups. In one preferred embodiment, an acetyl group is usedto protect the amino terminus and an amide group is used to protect thecarboxyl terminus. These blocking groups enhance the helix-formingtendencies of the peptides. Certain particularly preferred blockinggroups include alkyl groups of various lengths, e.g. groups having theformula: CH₃—(CH₂)_(n)—OO— where n ranges from about 1 to about 20,preferably from about 1 to about 16 or 18, more preferably from about 3to about 13, and most preferably from about 3 to about 10.

In certain particularly preferred embodiments, the protecting groupsinclude, but are not limited to alkyl chains as in fatty acids,propeonyl, formyl, and others. Particularly preferred carboxylprotecting groups include amides, esters, and ether-forming protectinggroups. In one preferred embodiment, an acetyl group is used to protectthe amino terminus and an amide group is used to protect the carboxylterminus. These blocking groups enhance the helix-forming tendencies ofthe peptides. Certain particularly preferred blocking groups includealkyl groups of various lengths, e.g. groups having the formula:CH₃—(CH₂)_(n)—OO— where n ranges from about 3 to about 20, preferablyfrom about 3 to about 16, more preferably from about 3 to about 13, andmost preferably from about 3 to about 10.

Other protecting groups include, but are not limited to Fmoc,t-butoxycarbonyl (t-BOC), 9-fluoreneacetyl group, 1-fluorenecarboxylicgroup, 9-florenecarboxylic group, 9-fluorenone-1-carboxylic group,benzyloxycarbonyl, Xanthyl (Xan), Trityl (Trt), 4-methyltrityl (Mtt),4-methoxytrityl (Mmt), 4-methoxy-2,3,6-trimethyl-benzenesulphonyl (Mtr),Mesitylene-2-sulphonyl (Mts), 4,4-dimethoxybenzhydryl (Mbh), Tosyl(Tos), 2,2,5,7,8-pentamethyl chroman-6-sulphonyl (Pmc), 4-methylbenzyl(MeBzl), 4-methoxybenzyl (MeOBzl), Benzyloxy (BzlO), Benzyl (Bzl),Benzoyl (Bz), 3-nitro-2-pyridinesulphenyl (Npys),1-(4,4-dimentyl-2,6-diaxocyclohexylidene)ethyl (Dde), 2,6-dichlorobenzyl(2,6-DiCl-Bzl), 2-chlorobenzyloxycarbonyl (2—Cl-Z),2-bromobenzyloxycarbonyl (2—Br-Z), Benzyloxymethyl (Born), cyclohexyloxy(cHxO),t-butoxymethyl (Bum), t-butoxy (tBuO), t-Butyl (tBu), Acetyl(Ac), and Trifluoroacetyl (TFA).

Protecting/blocking groups are well known to those of skill as aremethods of coupling such groups to the appropriate residue(s) comprisingthe peptides of this invention (see, e.g., Greene et al., (1991)Protective Groups in Organic Synthesis, 2nd ed., John Wiley & Sons, Inc.Somerset, N.J.). In one preferred embodiment, for example, acetylationis accomplished during the synthesis when the peptide is on the resinusing acetic anhydride. Amide protection can be achieved by theselection of a proper resin for the synthesis. During the synthesis ofthe peptides described herein in the examples, rink amide resin wasused. After the completion of the synthesis, the semipermanentprotecting groups on acidic bifunctional amino acids such as Asp and Gluand basic amino acid Lys, hydroxyl of Tyr are all simultaneouslyremoved. The peptides released from such a resin using acidic treatmentcomes out with the n-terminal protected as acetyl and the carboxylprotected as NH₂ and with the simultaneous removal of all of the otherprotecting groups.

In certain particularly preferred embodiments, the peptides comprise oneor more D-form (dextro rather than levo) amino acids as describedherein. In certain embodiments at least two enantiomeric amino acids,more preferably at least 4 enantiomeric amino acids and most preferablyat least 8 or 10 enantiomeric amino acids are “D” form amino acids. Incertain embodiments every other, or even every amino acid (e.g. everyenantiomeric amino acid) of the peptides described herein is a D-formamino acid.

In certain embodiments at least 50% of the enantiomeric amino acids are“D” form, more preferably at least 80% of the enantiomeric amino acidsare “D” form, and most preferably at least 90% or even all of theenantiomeric amino acids are “D” form amino acids.

G) Peptide Mimetics.

In addition to the peptides described herein, peptidomimetics are alsocontemplated. Peptide analogs are commonly used in the pharmaceuticalindustry as non-peptide drugs with properties analogous to those of thetemplate peptide. These types of non-peptide compound are termed“peptide mimetics” or “peptidomimetics” (Fauchere (1986) Adv. Drug Res.15: 29; Veber and Freidinger (1985) TINS p. 392; and Evans et al. (1987)J. Med. Chem. 30: 1229) and are usually developed with the aid ofcomputerized molecular modeling. Peptide mimetics that are structurallysimilar to therapeutically useful peptides may be used to produce anequivalent therapeutic or prophylactic effect.

Generally, peptidomimetics are structurally similar to a paradigmpolypeptide (e.g. SEQ ID NO:5 shown in Table 1), but have one or morepeptide linkages optionally replaced by a linkage selected from thegroup consisting of: —CH₂NH—, —CH₂—CH₂—, —CH═CH—(cis and trans),—COCH₂—, —CH(OH)CH₂—, —CH₂SO—, etc. by methods known in the art andfurther described in the following references: Spatola (1983) p. 267 inChemistry and Biochemistry of Amino Acids, Peptides, and Proteins, B.Weinstein, eds., Marcel Dekker, New York; Spatola (1983) Vega Data 1(3)Peptide Backbone Modifications. (general review); Morley (1980) TrendsPharm Sci pp. 463-468 (general review); Hudson et al. (1979) Int J PeptProt Res 14:177-185 (—CH₂NH—, CH₂CH₂—); Spatola et al. (1986) Life Sci38:1243-1249 (—CH₂—S); Harm, (1982) J Chem Soc Perkin Trans I 307-314(—CH—CH—, cis and trans); Almquist et al. (1980) J. Med. Chem.23:1392-1398 (—COCH₂—); Jennings-White et al. (1982) Tetrahedron Lett.23:2533 (—COCH₂—); Szelke et al., European Appln. EP 45665 (1982) CA:97:39405 (1982) (—CH(OH)CH₂—); Holladay et al. (1983) Tetrahedron Lett24:4401-4404 (—C(OH)CH₂—); and Hruby (1982) Life Sci., 31:189-199(—CH₂—S—)).

One particularly preferred non-peptide linkage is —CH₂NH—. Such peptidemimetics may have significant advantages over polypeptide embodiments,including, for example: more economical production, greater chemicalstability, enhanced pharmacological properties (half-life, absorption,potency, efficacy, etc.), reduced antigenicity, and others.

In addition, circularly permutations of the peptides described herein orconstrained peptides (including cyclized peptides) comprising aconsensus sequence or a substantially identical consensus sequencevariation may be generated by methods known in the art (Rizo andGierasch (1992) Ann. Rev. Biochem. 61: 387); for example, by addinginternal cysteine residues capable of forming intramolecular disulfidebridges which cyclize the peptide.

H) Small Organic Molecules.

In certain embodiments, the active agents of this invention includesmall organic molecules, e.g. as described in copending application U.S.Ser. No. 60/600,925, filed Aug. 11, 2004. In various embodiments thesmall organic molecules are similar to, and in certain cases, mimeticsof the tetra- and penta-peptides described in copending application U.S.Ser. No. 10/649,378, filed on Aug. 26, 2003 and U.S. Ser. No.60/494,449, filed on August 11.

The small organic molecules of this invention typically have molecularweights less than about 900 Daltons. Typically the molecules are highlysoluble in ethyl acetate (e.g., at concentrations equal to or greaterthan 4 mg/mL), and also are soluble in aqueous buffer at pH 7.0.

Contacting phospholipids such as1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), with the smallorganic molecules of this invention in an aqueous environment typicallyresults in the formation of particles with a diameter of approximately7.5 nm (.+-.0.1 nm). In addition, stacked bilayers are often foamed witha bilayer dimension on the order of 3.4 to 4.1 nm with spacing betweenthe bilayers in the stack of approximately 2 nm. Vesicular structures ofapproximately 38 nm are also often formed. Moreover, when the moleculesof this invention are administered to a mammal they render HDL moreanti-inflammatory and mitigate one or more symptoms of atherosclerosisand/or other conditions characterized by an inflammatory response.

Thus, in certain embodiments, the small organic molecule is one thatameliorates one or more symptoms of a pathology characterized by aninflammatory response in a mammal (e.g. atherosclerosis), where thesmall molecule is soluble in ethyl acetate at a concentration greaterthan 4 mg/mL, is soluble in aqueous buffer at pH 7.0, and, whencontacted with a phospholipid in an aqueous environment, forms particleswith a diameter of approximately 7.5 nm and forms stacked bilayers witha bilayer dimension on the order of 3.4 to 4.1 nm with spacing betweenthe bilayers in the stack of approximately 2 nm, and has a molecularweight less than 900 daltons.

In certain embodiment, the molecule has the formula:

where P¹, P², P³, and P⁴ are independently selected hydrophobicprotecting groups; R¹ and R⁴ are independently selected amino acid Rgroups; n, i, x, y, and z are independently zero or 1 such that when nand x are both zero, R¹ is a hydrophobic group and when y and i are bothzero, R⁴ is a hydrophobic group; R² and R³ are acidic or basic groups atpH 7.0 such that when R² is acidic, R³ is basic and when R² is basic, R³is acidic; and R⁵, when present is selected from the group consisting ofan aromatic group, an aliphatic group, a positively charged group, or anegatively charged group. In certain embodiments, R² or R³ is—(CH₂)_(j)—COOH where j=1, 2, 3, or 4 and/or —(CH₂)_(j)—NH₂ where j=1,2, 3, 4, or 5, or —(CH₂)_(j)—NH—C(═NH)—NH₂ where n=1, 2, 3 or 4. Incertain embodiments, R², R³, and R⁵, when present, are amino acid Rgroups. Thus, for example, In various embodiments R² and R³ areindependently an aspartic acid R group, a glutamic acid R group, alysine R group, a histidine R group, or an arginine R group (e.g., asillustrated in Table 1).

In certain embodiments, R¹ is selected from the group consisting of aLys R group, a Trp R group, a Phe R group, a Leu R group, an Orn Rgroup, pr a norLeu R group. In certain embodiments, R⁴ is selected fromthe group consisting of a Ser R group, a Thr R group, an Ile R group, aLeu R group, a norLeu R group, a Phe R group, or a Tyr R group.

In various embodiments x is 1, and R⁵ is an aromatic group (e.g., a TrpR group). In various embodiments at least one of n, x, y, and i is 1 andP¹, P², P³, and P⁴ when present, are independently selected from thegroup consisting of polyethylene glycol (PEG), an acetyl, amide, a 3 to20 carbon alkyl group, fmoc, 9-fluoreneacetyl group,1-fluorenecarboxylic group, 9-fluorenecarboxylic,9-fluorenone-1-carboxylic group, benzyloxycarbonyl, xanthyl (Xan),Trityl (Trt), 4-methyltrityl (Mtt), 4-methoxytrityl (Mmt),4-methoxy-2,3,6-trimethyl-benzenesulphonyl (Mtr), Mesitylene-2-sulphonyl(Mts), 4,4-dimethoxybenzhydryl (Mbh), Tosyl (Tos), 2,2,5,7,8-pentamethylchroman-6-sulphonyl (Pmc), 4-methylbenzyl (MeBzl), 4-methoxybenzyl(MeOBzl), benzyloxy (BzlO), benzyl (Bzl), benzoyl (Bz),3-nitro-2-pyridinesulphenyl (Npys),1-(4,4-dimethyl-2,6-dioxocyclohexylidene)ethyl (Dde), 2,6-dichlorobenzyl(2,6-DiCl-Bzl), 2-chlorobenzyloxycarbonyl (2—Cl-Z),2-bromobenzyloxycarbonyl (2—Br-Z), benzyloxymethyl (Born),t-butoxycarbonyl (Boc), cyclohexyloxy (cHxO), t-butoxymethyl (Bum),t-butoxy (tBuO), t-Butyl (tBu), a propyl group, a butyl group, a pentylgroup, a hexyl group, and trifluoroacetyl (TFA). In certain embodiments,P¹ when present and/or P² when present are independently selected fromthe group consisting of Boc-, Fmoc-, and Nicotinyl- and/or P³ whenpresent and/or P⁴ when present are independently selected from the groupconsisting of tBu, and OtBu.

While a number of protecting groups (P¹, P², P³, P⁴) are illustratedabove, this list is intended to be illustrative and not limiting. Inview of the teachings provided herein, a number of otherprotecting/blocking groups will also be known to one of skill in theart. Such blocking groups can be selected to minimize digestion (e.g.,for oral pharmaceutical delivery), and/or to increaseuptake/bioavailability (e.g., through mucosal surfaces in nasaldelivery, inhalation therapy, rectal administration), and/or to increaseserum/plasma half-life. In certain embodiments, the protecting groupscan be provided as an excipient or as a component of an excipient.

In certain embodiments, z is zero and the molecule has the formula:

where P¹, P², P³, P⁴, R¹, R², R³, R⁴, n, x, y, and i are as describedabove.

In certain embodiments, z is zero and the molecule has the formula:

where R¹, R², R³, and R⁴ are as described above.

In one embodiment, the molecule has the formula:

In certain embodiments, this invention contemplates small moleculeshaving one or more of the physical and/or functional propertiesdescribed herein and having the formula:

where P¹, P², P³, and P⁴ are independently selected hydrophobicprotecting groups as described above, n, x, and y are independently zeroor 1; j, k, and 1 are independently zero, 1, 2, 3, 4, or 5; and R² andR³ are acidic or basic groups at pH 7.0 such that when R² is acidic, R³is basic and when R² is basic, R³ is acidic. In certain preferredembodiments, the small molecule is soluble in water; and the smallmolecule has a molecular weight less than about 900 Daltons. In certainembodiments, n, x, y, j, and l are 1; and k is 4.

In certain embodiments, P¹ and/or P² are aromatic protecting groups. Incertain embodiments, R² and R³ are amino acid R groups, e.g., asdescribed above. In various embodiments least one of n, x, and y, is 1and P¹, P², P³ and P⁴ when present, are independently protecting groups,e.g. as described above. In certain embodiments the protecting groups,when present, are independently selected from the group consisting ofpolyethylene glycol (PEG), an acetyl, amide, 3 to 20 carbon alkylgroups, Fmoc, 9-fluoreneacetyl group, 1-fluorenecarboxylic group,9-fluorenecarboxylic, 9-fluorenone-1-carboxylic group,benzyloxycarbonyl, Xanthyl (Xan), Trityl (Trt), 4-methyltrityl (Mtt),4-methoxytrityl (Mmt), 4-methoxy-2,3,6-trimethyl-benzenesulphonyl (Mtr),Mesitylene-2-sulphonyl (Mts), 4,4-dimethoxybenzhydryl (Mbh), Tosyl(Tos), 2,2,5,7,8-penta.

III. Functional Assays of Active Agents

Certain active agents for use in the methods of this invention aredescribed herein by various formulas (e.g., Formula I, above) and/or byparticular sequences. In certain embodiments, preferred active agents ofthis invention are characterized by one or more of the followingfunctional properties:

1. They convert pro-inflammatory HDL to anti-inflammatory HDL or makeanti-inflammatory HDL more anti-inflammatory;

2. They decrease LDL-induced monocyte chemotactic activity generated byartery wall cells;

3. They stimulate the formation and cycling of pre-β HDL;

4. They raise HDL cholesterol; and/or

5. They increase HDL paraoxonase activity.

The specific agents disclosed herein, and/or agents corresponding to thevarious formulas described herein can readily be tested for one or moreof these activities as desired.

Methods of screening for each of these functional properties are wellknown to those of skill in the art. In particular, it is noted thatassays for monocyte chemotactic activity, HDL cholesterol, and HDLparaoxonase activity are illustrated in PCT/US01/26497 (WO 2002/15923).

IV. Peptide Preparation

The peptides used in this invention can be chemically synthesized usingstandard chemical peptide synthesis techniques or, particularly wherethe peptide does not comprise “D” amino acid residues, can berecombinantly expressed. In certain embodiments, even peptidescomprising “D” amino acid residues are recombinantly expressed. Wherethe polypeptides are recombinantly expressed, a host organism (e.g.bacteria, plant, fungal cells, etc.) in cultured in an environment whereone or more of the amino acids is provided to the organism exclusivelyin a D form. Recombinantly expressed peptides in such a system thenincorporate those D amino acids.

In preferred embodiments the peptides are chemically synthesized by anyof a number of fluid or solid phase peptide synthesis techniques knownto those of skill in the art. Solid phase synthesis in which theC-terminal amino acid of the sequence is attached to an insolublesupport followed by sequential addition of the remaining amino acids inthe sequence is a preferred method for the chemical synthesis of thepolypeptides of this invention. Techniques for solid phase synthesis arewell known to those of skill in the art and are described, for example,by Barany and Merrifield (1963) Solid-Phase Peptide Synthesis; pp. 3-284in The Peptides: Analysis, Synthesis, Biology. Vol. 2: Special Methodsin Peptide Synthesis, Part A.; Merrifield et al. (1963) J. Am. Chem.Soc., 85: 2149-2156, and Stewart et al. (1984) Solid Phase PeptideSynthesis, 2nd ed. Pierce Chem. Co., Rockford, Ill.

In certain embodiments, the peptides are synthesized by the solid phasepeptide synthesis procedure using a benzhyderylamine resin (BeckmanBioproducts, 0.59 mmol of NH₂/g of resin) as the solid support. The COOHterminal amino acid (e.g., t-butylcarbonyl-Phe) is attached to the solidsupport through a 4-(oxymethyl)phenacetyl group. This is a more stablelinkage than the conventional benzyl ester linkage, yet the finishedpeptide can still be cleaved by hydrogenation. Transfer hydrogenationusing formic acid as the hydrogen donor is used for this purpose.Detailed protocols used for peptide synthesis and analysis ofsynthesized peptides are described in a miniprint supplementaccompanying Anantharamaiah et al. (1985) J. Biol. Chem., 260(16):10248-10255.

It is noted that in the chemical synthesis of peptides, particularlypeptides comprising D amino acids, the synthesis usually produces anumber of truncated peptides in addition to the desired full-lengthproduct. The purification process (e.g. HPLC) typically results in theloss of a significant amount of the full-length product.

It was a discovery of this invention that, in the synthesis of a Dpeptide (e.g. D-4), in order to prevent loss in purifying the longestform one can dialyze and use the mixture and thereby eliminate the lastHPLC purification. Such a mixture loses about 50% of the potency of thehighly purified product (e.g. per wt of protein product), but themixture contains about 6 times more peptide and thus greater totalactivity.

V. Pharmaceutical Formulations and Devices A) PharmaceuticalFormulations

In order to carry out the methods of the invention, one or more activeagents of this invention are administered, e.g. to an individualdiagnosed as having one or more symptoms of atherosclerosis, or as beingat risk for atherosclerosis and or the various other pathologiesdescribed herein. The active agent(s) can be administered in the“native” form or, if desired, in the form of salts, esters, amides,prodrugs, derivatives, and the like, provided the salt, ester, amide,prodrug or derivative is suitable pharmacologically, i.e., effective inthe present method. Salts, esters, amides, prodrugs and otherderivatives of the active agents can be prepared using standardprocedures known to those skilled in the art of synthetic organicchemistry and described, for example, by March (1992) Advanced OrganicChemistry; Reactions, Mechanisms and Structure, 4th Ed. N.Y.Wiley-Interscience.

For example, acid addition salts are prepared from the free base usingconventional methodology, that typically involves reaction with asuitable acid. Generally, the base form of the drug is dissolved in apolar organic solvent such as methanol or ethanol and the acid is addedthereto. The resulting salt either precipitates or can be brought out ofsolution by addition of a less polar solvent. Suitable acids forpreparing acid addition salts include both organic acids, e.g., aceticacid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, malicacid, malonic acid, succinic acid, maleic acid, fumaric acid, tartaricacid, citric acid, benzoic acid, cinnamic acid, mandelic acid,methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid,salicylic acid, and the like, as well as inorganic acids, e.g.,hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid,phosphoric acid, and the like. An acid addition salt may be reconvertedto the free base by treatment with a suitable base. Particularlypreferred acid addition salts of the active agents herein are halidesalts, such as may be prepared using hydrochloric or hydrobromic acids.Conversely, preparation of basic salts of the active agents of thisinvention are prepared in a similar manner using a pharmaceuticallyacceptable base such as sodium hydroxide, potassium hydroxide, ammoniumhydroxide, calcium hydroxide, trimethylamine, or the like. Particularlypreferred basic salts include alkali metal salts, e.g., the sodium salt,and copper salts.

Preparation of Esters Typically Involves Functionalization of Hydroxyland/or carboxyl groups which may be present within the molecularstructure of the drug. The esters are typically acyl-substitutedderivatives of free alcohol groups, i.e., moieties that are derived fromcarboxylic acids of the formula RCOOH where R is alky, and preferably islower alkyl. Esters can be reconverted to the free acids, if desired, byusing conventional hydrogenolysis or hydrolysis procedures.

Amides and prodrugs can also be prepared using techniques known to thoseskilled in the art or described in the pertinent literature. Forexample, amides may be prepared from esters, using suitable aminereactants, or they may be prepared from an anhydride or an acid chlorideby reaction with ammonia or a lower alkyl amine. Prodrugs are typicallyprepared by covalent attachment of a moiety that results in a compoundthat is therapeutically inactive until modified by an individual'smetabolic system.

The active agents identified herein are useful for parenteral, topical,oral, nasal (or otherwise inhaled), rectal, or local administration,such as by aerosol or transdermally, for prophylactic and/or therapeutictreatment of one or more of the pathologies/indications described herein(e.g., atherosclerosis and/or eye disease and/or symptoms thereof). Thepharmaceutical compositions can be administered in a variety of unitdosage forms depending upon the method of administration. Suitable unitdosage forms, include, but are not limited to powders, tablets, pills,capsules, lozenges, suppositories, patches, nasal sprays, injectibles,implantable sustained-release formulations, lipid complexes, etc.

The active agents of this invention are typically combined with apharmaceutically acceptable carrier (excipient) to form apharmacological composition. Pharmaceutically acceptable carriers cancontain one or more physiologically acceptable compound(s) that act, forexample, to stabilize the composition or to increase or decrease theabsorption of the active agent(s). Physiologically acceptable compoundscan include, for example, carbohydrates, such as glucose, sucrose, ordextrans, antioxidants, such as ascorbic acid or glutathione, chelatingagents, low molecular weight proteins, protection and uptake enhancerssuch as lipids, compositions that reduce the clearance or hydrolysis ofthe active agents, or excipients or other stabilizers and/or buffers.

Other physiologically acceptable compounds include wetting agents,emulsifying agents, dispersing agents or preservatives that areparticularly useful for preventing the growth or action ofmicroorganisms. Various preservatives are well known and include, forexample, phenol and ascorbic acid. One skilled in the art wouldappreciate that the choice of pharmaceutically acceptable carrier(s),including a physiologically acceptable compound depends, for example, onthe route of administration of the active agent(s) and on the particularphysio-chemical characteristics of the active agent(s).

The excipients are preferably sterile and generally free of undesirablematter. These compositions may be sterilized by conventional, well-knownsterilization techniques.

In therapeutic applications, the compositions of this invention areadministered to a patient suffering from one or more symptoms of the oneor more pathologies described herein, or at risk for one or more of thepathologies described herein in an amount sufficient to prevent and/orcure and/or or at least partially prevent or arrest the disease and/orits complications. An amount adequate to accomplish this is defined as a“therapeutically effective dose.” Amounts effective for this use willdepend upon the severity of the disease and the general state of thepatient's health. Single or multiple administrations of the compositionsmay be administered depending on the dosage and frequency as requiredand tolerated by the patient. In any event, the composition shouldprovide a sufficient quantity of the active agents of the formulationsof this invention to effectively treat (ameliorate one or more symptoms)the patient.

The concentration of active agent(s) can vary widely, and will beselected primarily based on fluid volumes, viscosities, body weight andthe like in accordance with the particular mode of administrationselected and the patient's needs. Concentrations, however, willtypically be selected to provide dosages ranging from about 0.1 or 1mg/kg/day to about 50 mg/kg/day and sometimes higher. Typical dosagesrange from about 3 mg/kg/day to about 3.5 mg/kg/day, preferably fromabout 3.5 mg/kg/day to about 7.2 mg/kg/day, more preferably from about7.2 mg/kg/day to about 11.0 mg/kg/day, and most preferably from about11.0 mg/kg/day to about 15.0 mg/kg/day. In certain preferredembodiments, dosages range from about 10 mg/kg/day to about 50mg/kg/day. In certain embodiments, dosages range from about 20 mg toabout 50 mg given orally twice daily. It will be appreciated that suchdosages may be varied to optimize a therapeutic regimen in a particularsubject or group of subjects. For example, the concentration fortreating an eye disease can be selected to provide dosages ranging from200 ug/ml to 800 ug/ml of fluid.

In certain preferred embodiments, the active agents of this inventionare administered orally (e.g. via a tablet) or as an injectable inaccordance with standard methods well known to those of skill in theart. In other preferred embodiments, the peptides, may also be deliveredthrough the skin using conventional transdermal drug delivery systems,i.e., transdermal “patches” wherein the active agent(s) are typicallycontained within a laminated structure that serves as a drug deliverydevice to be affixed to the skin. In such a structure, the drugcomposition is typically contained in a layer, or “reservoir,”underlying an upper backing layer. It will be appreciated that the term“reservoir” in this context refers to a quantity of “activeingredient(s)” that is ultimately available for delivery to the surfaceof the skin. Thus, for example, the “reservoir” may include the activeingredient(s) in an adhesive on a backing layer of the patch, or in anyof a variety of different matrix formulations known to those of skill inthe art. The patch may contain a single reservoir, or it may containmultiple reservoirs.

In one embodiment, the reservoir comprises a polymeric matrix of apharmaceutically acceptable contact adhesive material that serves toaffix the system to the skin during drug delivery. Examples of suitableskin contact adhesive materials include, but are not limited to,polyethylenes, polysiloxanes, polyisobutylenes, polyacrylates,polyurethanes, and the like. Alternatively, the drug-containingreservoir and skin contact adhesive are present as separate and distinctlayers, with the adhesive underlying the reservoir which, in this case,may be either a polymeric matrix as described above, or it may be aliquid or hydrogel reservoir, or may take some other form. The backinglayer in these laminates, which serves as the upper surface of thedevice, preferably functions as a primary structural element of the“patch” and provides the device with much of its flexibility. Thematerial selected for the backing layer is preferably substantiallyimpermeable to the active agent(s) and any other materials that arepresent.

Other preferred formulations for topical drug delivery include, but arenot limited to, ointments and creams. Ointments are semisolidpreparations which are typically based on petrolatum or other petroleumderivatives. Creams containing the selected active agent, are typicallyviscous liquid or semisolid emulsions, often either oil-in-water orwater-in-oil. Cream bases are typically water-washable, and contain anoil phase, an emulsifier and an aqueous phase. The oil phase, alsosometimes called the “internal” phase, is generally comprised ofpetrolatum and a fatty alcohol such as cetyl or stearyl alcohol; theaqueous phase usually, although not necessarily, exceeds the oil phasein volume, and generally contains a humectant. The emulsifier in a creamformulation is generally a nonionic, anionic, cationic or amphotericsurfactant. The specific ointment or cream base to be used, as will beappreciated by those skilled in the art, is one that will provide foroptimum drug delivery. As with other carriers or vehicles, an ointmentbase should be inert, stable, nonirritating and nonsensitizing.

In addition, the active agents of this invention can be administered viaintraocular injection (e.g. intravitreal injection) in accordance withstandard methods well known to those of skill in the art.

Unlike typical peptide formulations, the peptides of this inventioncomprising D-form amino acids can be administered, even orally, withoutprotection against proteolysis by stomach acid, etc. Nevertheless, incertain embodiments, peptide delivery can be enhanced by the use ofprotective excipients. This is typically accomplished either bycomplexing the polypeptide with a composition to render it resistant toacidic and enzymatic hydrolysis or by packaging the polypeptide in anappropriately resistant carrier such as a liposome. Means of protectingpolypeptides for oral delivery are well known in the art (see, e.g.,U.S. Pat. No. 5,391,377 describing lipid compositions for oral deliveryof therapeutic agents).

Elevated serum half-life can be maintained by the use ofsustained-release protein “packaging” systems. Such sustained releasesystems are well known to those of skill in the art. In one preferredembodiment, the ProLease biodegradable microsphere delivery system forproteins and peptides (Tracy (1998) Biotechnol. Prog., 14: 108; Johnsonet al. (1996) Nature Med. 2: 795; Herbert et al. (1998), Pharmaceut.Res. 15, 357) a dry powder composed of biodegradable polymericmicrospheres containing the active agent in a polymer matrix that can becompounded as a dry formulation with or without other agents.

The ProLease microsphere fabrication process was specifically designedto achieve a high encapsulation efficiency while maintaining integrityof the active agent. The process consists of (i) preparation offreeze-dried drug particles from bulk by spray freeze-drying the drugsolution with stabilizing excipients, (ii) preparation of a drug-polymersuspension followed by sonication or homogenization to reduce the drugparticle size, (iii) production of frozen drug-polymer microspheres byatomization into liquid nitrogen, (iv) extraction of the polymer solventwith ethanol, and (v) filtration and vacuum drying to produce the finaldry-powder product. The resulting powder contains the solid form of theactive agents, which is homogeneously and rigidly dispersed withinporous polymer particles. The polymer most commonly used in the process,poly(lactide-co-glycolide) (PLG), is both biocompatible andbiodegradable.

Encapsulation can be achieved at low temperatures (e.g., −40° C.).During encapsulation, the protein is maintained in the solid state inthe absence of water, thus minimizing water-induced conformationalmobility of the protein, preventing protein degradation reactions thatinclude water as a reactant, and avoiding organic-aqueous interfaceswhere proteins may undergo denaturation. A preferred process usessolvents in which most proteins are insoluble, thus yielding highencapsulation efficiencies (e.g., greater than 95%).

In another embodiment, one or more components of the solution can beprovided as a “concentrate”, e.g., in a storage container (e.g., in apremeasured volume) ready for dilution, or in a soluble capsule readyfor addition to a volume of water.

The foregoing formulations and administration methods are intended to beillustrative and not limiting. It will be appreciated that, using theteaching provided herein, other suitable formulations and modes ofadministration can be readily devised.

B) Lipid-Based Formulations

In certain embodiments, the active agents of this invention areadministered in conjunction with one or more lipids. The lipids can beformulated as an excipient to protect and/or enhance transport/uptake ofthe active agents or they can be administered separately.

Without being bound by a particular theory, it was discovered of thisinvention that administration (e.g. oral administration) of certainphospholipids can significantly increase HDL/LDL ratios. In addition, itis believed that certain medium-length phospholipids are transported bya process different than that involved in general lipid transport. Thus,co-administration of certain medium-length phospholipids with the activeagents of this invention confer a number of advantages: They protect theactive agents from digestion or hydrolysis, they improve uptake, andthey improve HDL/LDL ratios.

The lipids can be formed into liposomes that encapsulate the activeagents of this invention and/or they can be complexed/admixed with theactive agents and/or they can be covalently coupled to the activeagents. Methods of making liposomes and encapsulating reagents are wellknown to those of skill in the art (see, e.g., Martin andPapahadjopoulos (1982) J. Biol. Chem., 257: 286-288; Papahadjopoulos etal. (1991) Proc. Natl. Acad. Sci. USA, 88: 11460-11464; Huang et al.(1992) Cancer Res., 52:6774-6781; Lasic et al. (1992) FEBS Lett., 312:255-258., and the like).

Preferred phospholipids for use in these methods have fatty acidsranging from about 4 carbons to about 24 carbons in the sn-1 and sn-2positions. In certain preferred embodiments, the fatty acids aresaturated. In other preferred embodiments, the fatty acids can beunsaturated. Various preferred fatty acids are illustrated in Table 19.

TABLE 19 Suitable fatty acids in the sn-1 and/or sn-2 position of thepreferred phospholipids for administration of active agents describedherein. Carbon No. Common Name IUPAC Name  3:0 Propionoyl Trianoic  4:0Butanoyl Tetranoic  5:0 Pentanoyl Pentanoic  6:0 Caproyl Hexanoic  7:0Heptanoyl Heptanoic  8:0 Capryloyl Octanoic  9:0 Nonanoyl Nonanoic 10:0Capryl Decanoic 11:0 Undcanoyl Undecanoic 12:0 Lauroyl Dodecanoic 13:0Tridecanoyl Tridecanoic 14:0 Myristoyl Tetradecanoic 15:0 PentadecanoylPentadecanoic 16:0 Palmitoyl Hexadecanoic 17:0 HeptadecanoylHeptadecanoic 18:0 Stearoyl Octadecanoic 19:0 Nonadecanoyl Nonadecanoic20:0 Arachidoyl Eicosanoic 21:0 Heniecosanoyl Heniecosanoic 22:0Behenoyl Docosanoic 23:0 Trucisanoyl Trocosanoic 24:0 LignoceroylTetracosanoic 14:1 Myristoleoyl (9-cis) 14:1 Myristelaidoyl (9-trans)16:1 Palmitoleoyl (9-cis) 16:1 Palmitelaidoyl (9-trans)

The fatty acids in these positions can be the same or different.Particularly preferred phospholipids have phosphorylcholine at the sn-3position.

C) Specialized Delivery/Devices 1. Drug-Eluting Stents

Restenosis, the reclosure of a previously stenosed and subsequentlydilated peripheral or coronary vessel occurs at a significant rate(e.g., 20-50% for these procedures) and is dependent on a number ofclinical and morphological variables. Restenosis may begin shortlyfollowing an angioplasty procedure, but usually ceases at the end ofapproximately six (6) months.

A recent technology that has been developed to address the problem ofrestenosis in intravascular stents. Stents are typically devices thatare permanently implanted (expanded) in coronary and peripheral vessels.The goal of these stents is to provide a long-term “scaffolding” orsupport for the diseased (stenosed) vessels. The theory being, if thevessel is supported from the inside, it will not close down orrestenose.

Known stent designs include, but are not limited to monofilament wirecoil stents (see, e.g., U.S. Pat. No. 4,969,458 which is incorporatedherein by reference); welded metal cages (see, e.g., U.S. Pat. Nos.4,733,665 and 4,776,337 which are incorporated herein by reference),thin-walled metal cylinders with axial slots formed around thecircumference (see, e.g., U.S. Pat. Nos. 4,733,665, 4,739,762, 4,776,337which are incorporated herein by reference). Known constructionmaterials for use in stents include, but are not limited to polymers,organic fabrics and biocompatible metals, such as, stainless steel,gold, silver, tantalum, titanium, and shape memory alloys such asNitinol.

To further prevent restenosis, stents can be covered and/or impregnatedwith one or more pharmaceutical, e.g., in controlled releaseformulations to inhibit cell proliferation associated with restenosis.Most commonly such “drug-eluting” stents are designed to deliver variouscancer drugs (cytotoxins).

However, because of their activity in mitigating inflammatory responses,reducing and/or eliminated oxidized lipids and/or other oxidizedspecies, inhibiting macrophage chemotactic activity and the like, theactive agents described herein are well suited to prevent restenosis.Thus, in certain embodiments, this invention contemplates stents havingone or more of the active agents described herein coated on the surfaceand/or retained within cavities or microcavities in the surface of thestent.

In certain embodiments the active agents are contained withinbiocompatible matrices (e.g. biocompatible polymers such as urethane,silicone, and the like). Suitable biocompatible materials are described,for example, in U.S. Patent Publications 2005/0084515, 2005/00791991,2005/0070996, and the like which are incorporated herein by reference.In various embodiments the polymers include, but are not limited tosilicone-urethane copolymer, a polyurethane, a phenoxy, ethylene vinylacetate, polycaprolactone, poly(lactide-co-glycolide), polylactide,polysulfone, elastin, fibrin, collagen, chondroitin sulfate, abiocompatible polymer, a biostable polymer, a biodegradable polymer

Thus, in certain embodiments this invention provides a stent fordelivering drugs to a vessel in a body. The stent typically comprisesstent framework including a plurality of reservoirs formed therein. Thereservoirs typically include an active agent and/or activeagent-containing polymer positioned in the reservoir and/or coated onthe surface of the stent. In various embodiments the stent is a metallicbase or a polymeric base. Certain preferred stent materials include, butare not limited to stainless steel, nitinol, tantalum, MP35N alloy,platinum, titanium, a suitable biocompatible alloy, a suitablebiocompatible polymer, and/or a combination thereof.

In various embodiments where the stent comprises pores (e.g.reservoirs), the pores can include micropores (e.g., having a diameterthat ranges from about 10 to about 50 μm, preferably about 20 μm orless). In various embodiments the micropores have a depth in the rangeof about 10 μm to about 50 μm. In various embodiments the microporesextend through the stent framework having an opening on an interiorsurface of the stent and an opening on an exterior surface of the stent.In certain embodiments the stent can, optionally comprise a cap layerdisposed on the interior surface of the stent framework, the cap layercovering at least a portion of the through-holes and providing a barriercharacteristic to control an elution rate of the active agent(s) in thepolymer from the interior surface of the stent framework. In variousembodiments the reservoirs comprise channels along an exterior surfaceof the stent framework. The stent can optionally have multiple layers ofpolymer where different layers of polymer carry different activeagent(s) and/or other drugs.

In certain embodiments the stent comprises: an adhesion layer positionedbetween the stent framework and the polymer. Suitable adhesion layersinclude, but are not limited to a polyurethane, a phenoxy,poly(lactide-co-glycolide)-, polylactide, polysulfone, polycaprolactone,an adhesion promoter, and/or a combination thereof.

In addition to stents, the active agents can be coated on or containedwithin essentially any implantable medical device configured forimplantation in a extravascular and/or intravascular location.

Also provided are methods of manufacturing a drug-polymer stent,comprising. The methods involve providing a stent framework; cutting aplurality of reservoirs in the stent framework, e.g., using a high powerlaser; applying one or more of the active agents and/or a drug polymerto at least one reservoir; drying the drug polymer; applying a polymerlayer to the dried drug polymer; and drying the polymer layer. Theactive agent(s) and/or polymer(s) can be applied by any convenientmethod including but not limited to spraying, dipping, painting,brushing and dispensing.

Also provided are methods of treating a vascular condition and/or acondition characterized by an inflammatory response and/or a conditioncharacterized by the formation of oxidized reactive species. The methodstypically involve positioning a stent or other implantable device asdescribed above within the body (e.g. within a vessel of a body) andeluting at least active agent from at least one surface of the implant.

2. Impregnated Grafts and Transplants

Vascular grafts can be classified as either biological or synthetic.There are two commonly used types of biological grafts. An autograft isone taken from another site in the patient. In peripheral vascularsurgery by far the most commonly used such graft is the long saphenousvein. This can be used in situ with the valves surgically destroyed withan intraluminal cutting valvutome.

Alternatively, the vein can be removed and reversed but this typicallyproduces a discrepancy between the anastomotic size of the artery andvein. In thoracic surgery the use of internal mammary artery forcoronary artery bypass surgery is another example of an autograft. Anallograft is one taken from another animal of the same species.Externally supported umbilical vein is rarely used but is an example ofsuch a graft.

Synthetic grafts are most commonly made from Dacron orpolytetrafluoroethylene (PTFE). Dacron grafts are frequently used inaortic and aorto-iliac surgery. Below the inguinal ligament the resultsof all synthetic grafts are inferior to those obtained with the use ofvein grafts. Suitable vein is not always available and in this situationPTFE is typically used. It can be used in conjunction with vein as acomposite graft. Neointimal hyperplasia at the distal anastomosis can bereduced by the incorporation of a segment of vein as either a MillarCuff or Taylor Patch to improve the long-term patency of the grafts.

The commonest complications associated with the use of vascular graftsinclude Graft occlusion, Graft infection, true and false aneurysms atthe site of anastomosis, distal embolization, and erosion in to adjacentstructures—e.g. Aorto-enteric fistulae. Many of these conditions areassociated with an inflammatory response, macrophage migration into thesite, and/or the formation of reactive oxygen species (e.g., oxidizedlipids). To reduce such complications, the graft (synthetic orbiological can be soaked, or otherwise coated, with one or more of theactive agents described herein.

In addition, it is contemplated that other implantable tissues ormaterials can similarly be impregnated or coated with one or more activeagents of this invention. Thus, for example, in certain embodiments thisinvention contemplates the use of impregnated sutures to minimizeinflammation and/or infection and/or tissue rejection.

3. Subcutaneous Matrices

In certain embodiments, one or more active agents described herein areadministered alone or in combination with other therapeutics asdescribed herein in implantable (e.g., subcutaneous) matrices.

A major problem with standard drug dosing is that typical delivery ofdrugs results in a quick burst of medication at the time of dosing,followed by a rapid loss of the drug from the body. Most of the sideeffects of a drug occur during the burst phase of its release into thebloodstream. Secondly, the time the drug is in the bloodstream attherapeutic levels is very short, most is used and cleared during theshort burst.

Drugs (e.g., the active agents described herein) imbedded in variousmatrix materials for sustained release provides some solution to theseproblems. Drugs embedded, for example, in polymer beads or in polymerwafers have several advantages. First, most systems allow slow releaseof the drug, thus creating a continuous dosing of the body with smalllevels of drug. This typically prevents side effects associated withhigh burst levels of normal injected or pill based drugs. Secondly,since these polymers can be made to release over hours to months, thetherapeutic span of the drug is markedly increased. Often, by mixingdifferent ratios of the same polymer components, polymers of differentdegradation rates can be made, allowing remarkable flexibility dependingon the agent being used. A long rate of drug release is beneficial forpeople who might have trouble staying on regular dosage, such as theelderly, but is also an ease of use improvement that everyone canappreciate. Most polymers can be made to degrade and be cleared by thebody over time, so they will not remain in the body after thetherapeutic interval.

Another advantage of polymer based drug delivery is that the polymersoften can stabilize or solubilize proteins, peptides, and other largemolecules that would otherwise be unusable as medications. Finally, manydrug/polymer mixes can be placed directly in the disease area, allowingspecific targeting of the medication where it is needed without losingdrug to the “first pass” effect. This is certainly effective fortreating the brain, which is often deprived of medicines that can'tpenetrate the blood/brain barrier.

A number of implantable matrix (sustained release) systems are know tothose of skill and can readily be adapted for use with one or more ofthe active agents described herein. Suitable sustained release systemsinclude, but are not limited to Re-Gel®, SQ2Gel®, and Oligosphere® byMacroMed, ProLease® and Medisorb® by Alkermes, Paclimer® and Gliadel®Wafer by Guilford pharmaceuticals, the Duros implant by Alza, acousticbioSpheres by Point Biomedical, the Intelsite capsule by Scintipharma,Inc., and the like.

4. Other “Specialty Delivery Systems”

Other “specialty” delivery systems include, but are not limited to lipidbased oral mist that allows absorption of drugs across the oral mucosa,developed by Generex Biotechnology, the oral transmucosal system (OTS™)by Anesta Corp., the inhalable dry powder and PulmoSpheres technology byInhale Therapeutics, the AERx® Pulmonary Drug Delivery System byAradigm, the AIR mechanism by Alkermes, and the like.

Another approach to delivery developed by Alkermes is a system targetedfor elderly and pediatric use, two populations for which taking pills isoften difficult is known as Drug Sipping Technology (DST). Themedication is placed in a drinking straw device, prevented from fallingout by filters on either end of it. The patient merely has to drinkclear liquid (water, juice, soda) through the straw. The drug dissolvesin the liquid as it is pulled through and is ingested by the patient.The filter rises to the top of the straw when all of the medication istaken. This method has the advantage in that it is easy to use, theliquid often masks the medication's taste, and the drug is pre-dissolvedfor more efficient absorption.

It is noted that these uses and delivery systems are intended to beillustrative and not limiting. Using the teachings provided herein,other uses and delivery systems will be known to those of skill in theart.

VI. Additional Pharmacologically Active Agents Combined Active Agents

In various embodiments, the use of combinations of two or more activeagents described is contemplated in the treatment of the variouspathologies/indications described herein. The use of combinations ofactive agents can alter pharmacological activity, bioavailability, andthe like.

By way of illustration, it is noted that D-4F rapidly associates withpre-beta HDL and HDL and then is rapidly cleared from the circulation(it is essentially non-detectable 6 hours after an oral dose), whileD-[113-122]apoJ slowly associates with pre-beta HDL and to a lesserextent with HDL but remains associated with these HDL fractions for atleast 36 hours. FREL associates with HDL and only HDL but remainsdetectable in HDL for much longer than D-4F (i.e., it is detectable inHDL 48 hours after a single oral dose in mice). In certain embodimentsthis invention thus contemplates combinations of, for example, thesethree peptides to reduce the amount to reduce production expense, and/orto optimize dosage regimen, therapeutic profile, and the like. Incertain embodiments combinations of the active agents described hereincan be simply coadministered and/or added together to form a singlepharmaceutical formulation. In certain embodiments the various activeagent(s) can be complexed together (e.g. via hydrogen bonding) to formactive agent complexes that are more effective than the parent agents.

Use with Additional Pharmacologically Active Materials.

Additional pharmacologically active materials (i.e., drugs) can bedelivered in conjunction with one or more of the active agents describedherein. In certain embodiments, such agents include, but are not limitedto agents that reduce the risk of atherosclerotic events and/orcomplications thereof. Such agents include, but are not limited to betablockers, beta blockers and thiazide diuretic combinations, statins,aspirin, ace inhibitors, ace receptor inhibitors (ARBs), and the like.

It was discovered that, adding a low dosage active agent (e.g., of D-4F)(1 μg/ml) to the drinking water of apoE null mice for 24 hours did notsignificantly improve HDL function (see, e.g., related application U.S.Ser. No. 10/423,830, filed on Apr. 25, 2003, which is incorporatedherein by reference). In addition, adding 0.05 mg/ml of atorvastatin orpravastatin alone to the drinking water of the apoE null mice for 24hours did not improve HDL function. However, when D-4F 1 μg/ml was addedto the drinking water together with 0.05 mg/ml of atorvastatin orpravastatin there was a significant improvement in HDL function). Indeedthe pro-inflammatory apoE null HDL became as anti-inflammatory as 350μg/ml of normal human HDL (h, HDL see, e.g., related application U.S.Ser. No. 10/423,830).

Thus, doses of D-4F alone, or statins alone, which by themselves had noeffect on HDL function when given together acted synergistically. WhenD-4F and a statin were given together to apo E null mice, theirpro-inflammatory HDL at 50 μg/ml of HDL-cholesterol became as effectiveas normal human HDL at 350 μg/ml of HDL-cholesterol in preventing theinflammatory response induced by the action of HPODE oxidizing PAPC incocultures of human artery wall cells.

Thus, in certain embodiments this invention provides methods forenhancing the activity of statins. The methods generally involveadministering one or more of the active agents described herein, asdescribed herein in conjunction with one or more statins. The activeagents achieve synergistic action between the statin and the agent(s) toameliorate one or more symptoms of atherosclerosis. In this contextstatins can be administered at significantly lower dosages therebyavoiding various harmful side effects (e.g., muscle wasting) associatedwith high dosage statin use and/or the anti-inflammatory properties ofstatins at any given dose are significantly enhanced.

Suitable statins include, but are not limited to pravastatin(Pravachol/Bristol-Myers Squibb), simvastatin (Zocor/Merck), lovastatin(Mevacor/Merck), and the like.

In various embodiments the active agent(s) described herein areadministered in conjunction with one or more beta blockers. Suitablebeta blockers include, but are not limited to cardioselective (selectivebeta 1 blockers), e.g., acebutolol (Sectral™), atenolol (Tenormin™),betaxolol (Kerlone™), bisoprolol (Zebeta™), metoprolol (Lopressor™), andthe like. Suitable non-selective blockers (block beta 1 and beta 2equally) include, but are not limited to carteolol (Cartrol™), nadolol(Corgard™), penbutolol (Levatol™), pindolol (Visken™), propranolol(Inderal™), timolol (Blockadren™), labetalol (Normodyne™, Trandate™),and the like.

Suitable beta blocker thiazide diuretic combinations include, but arenot limited to Lopressor HCT, ZIAC, Tenoretic, Corzide, Timolide,Inderal LA 40/25, Inderide, Normozide, and the like.

Suitable ace inhibitors include, but are not limited to captopril (e.g.Capoten™ by Squibb), benazepril (e.g., Lotensin™ by Novartis), enalapril(e.g., Vasotec™ by Merck), fosinopril (e.g., Monopril™ byBristol-Myers), lisinopril (e.g. Prinivil™ by Merck or Zestril™ byAstra-Zeneca), quinapril (e.g. Accupril™ by Parke-Davis), ramipril(e.g., Altace™ by Hoechst Marion Roussel, King Pharmaceuticals),imidapril, perindopril erbumine (e.g., Aceon™ by Rhone-Polenc Rorer),trandolapril (e.g., Mavik™ by Knoll Pharmaceutical), and the like.Suitable ARBS (Ace Receptor Blockers) include but are not limited tolosartan (e.g. Cozaar™ by Merck), irbesartan (e.g., Avapro™ by Sanofi),candesartan (e.g., Atacand™ by Astra Merck), valsartan (e.g., Diovan™ byNovartis), and the like.

In various embodiments, one or more agents described herein areadministered with one or more of the drugs identified below.

Thus, in certain embodiments one or more active agents are administeredin conjunction with cholesteryl ester transfer protein (CETP) inhibitors(e.g., torcetrapib, JTT-705. CP-529414) and/or acyl-CoA:cholesterolO-acyltransferase (ACAT) inhibitors (e.g., Avasimibe (CI-1011), CP113818, F-1394, and the like), and/or immunomodulators (e.g., FTY720(sphingosine-1-phosphate receptor agonist), Thalomid (thalidomide),Imuran (azathioprine), Copaxone (glatiramer acetate), Certican®(everolimus), Neoral® (cyclosporine), and the like), and/ordipeptidyl-peptidase-4 (DPP4) inhibitors (e.g.,2-Pyrrolidinecarbonitrile,1-[[[2-[(5-cyano-2-pyridinyl)amino]ethyl]amino]acetyl], see also U.S.Patent Publication 2005-0070530), and/or calcium channel blockers (e.g.,Adalat, Adalat CC, Calan, Calan SR, Cardene, Cardizem, Cardizem CD,Cardizem SR, Dilacor-XR, DynaCirc, Isoptin, Isoptin SR, Nimotop,Norvasc, Plendil, Procardia, Procardia XL, Vascor, Verelan), and/orperoxisome proliferator-activated receptor (PPAR) agonists for, e.g., α,γ, receptors (e.g., Azelaoyl PAF, 2-Bromohekadecanoic acid, Ciglitizone,Clofibrate, 15-Deoxy-δ¹2,14-prostaglandin J₂, Fenofibrate, Fmoc-Leu-OH,GW1929, GW7647, 8(S)-Hydroxy-(5Z,9E,11Z,14Z)-eicosatetraenoic acid(8(S)-HETE), Leukotriene B₄, LY-171,883 (Tomelukast), Prostaglandin A₂,Prostaglandin J₂, Tetradecylthioacetic acid (TTA), Troglitazone(CS-045), WY-14643 (Pirinixic acid)), and the like.

In certain embodiments one or more of the active agents are administeredin conjunction with fibrates (e.g., clofibrate (atromid), gemfibrozil(lopid), fenofibrate (tricor), etc.), bile acid sequestrants (e.g.,cholestyramine, colestipol, etc.), cholesterol absorption blockers(e.g., ezetimibe (Zetia), etc.), Vytorin ((ezetimibe/simvastatincombination), and/or steroids, warfarin, and/or aspirin, and/or Bcr-Ablinhibitors/antagonists (e.g., Gleevec (Imatinib Mesylate), AMN₁O₇,STI571 (CGP57148B), ON 012380, PLX225, and the like), and/or reninangiotensin pathway blockers (e.g., Losartan (Cozaar®), Valsartan(Diovan®), Irbesartan (Avapro®), Candesartan (Atacand®), and the like),and/or angiotensin II receptor antagonists (e.g., losartan (Cozaar),valsartan (Diovan), irbesartan (Avapro), candesartan (Atacand) andtelmisartan (Micardis), etc.), and/or PKC inhibitors (e.g., CalphostinC, Chelerythrine chloride, Chelerythrine.chloride, Copperbis-3,5-diisopropylsalicylate, Ebselen, EGF Receptor (human) (651-658)(N-Myristoylated), Go 6976, H-7.dihydrochloride,1-O-Hexadecyl-2-O-methyl-rac-glycerol, Hexadecyl-phosphocholine(C_(16:0)); Miltefosine, Hypericin, Melittin (natural), Melittin(synthetic), ML-7.hydrochloride, ML-9.hydrochloride,Palmitoyl-DL-carnitine.hydrochloride, Protein Kinase C (19-31), ProteinKinase C (19-36), Quercetin.dihydrate, Quercetin.dihydrate,D-erythro-Sphingosine (isolated), D-erythro-Sphingosine (synthetic),Sphingosine, N,N-dimethyl, D-erythro-Sphingosine, Dihydro-,D-erythro-Sphingosine, N,N-Dimethyl-, D-erythro-Sphingosine chloride,N,N,N-Trimethyl-, Staurosporine, Bisindolylmaleimide I, G-6203, and thelike).

In certain embodiments, one or more of the active agents areadministered in conjunction with ApoAI, Apo A-I derivatives and/oragonists (e.g., ApoAI milano, see, e.g., U.S. Patent Publications20050004082, 20040224011, 20040198662, 20040181034, 20040122091,20040082548, 20040029807, 20030149094, 20030125559, 20030109442,20030065195, 20030008827, and 20020071862, and U.S. Pat. Nos. 6,831,105,6,790,953, 6,773,719, 6,713,507, 6,703,422, 6,699,910, 6,680,203,6,673,780, 6,646,170, 6,617,134, 6,559,284, 6,506,879, 6,506,799,6,459,003, 6,423,830, 6,410,802, 6,376,464, 6,367,479, 6,329,341,6,287,590, 6,090,921, 5,990,081, and the like), renin inhibitors (e.g.,SPP630 and SPP635, SPP100, Aliskiren, and the like), and/or MRantagonist (e.g., spironolactone, aldosterone glucuronide, and thelike), and/or aldosterone synthase inhibitors, and/or alpha-adrenergicantagonists (e.g., Aldomet® (Methyldopa), Cardura® (Doxazocin),Catapres®; Catapres-TTS®; Duraclon™ (Clonidine), Dibenzyline®(Phenoxybenzamine), Hylorel® (Guanadrel), Hytrin® (Terazosin),Minipress® (Prazosin), Tenex® (Guanfacine), Guanabenz, Phentolamine,Reserpine, and the like), and/or liver X receptor (LXR) agonists (e.g.,T0901317, GW3965, ATI-829, acetyl-podocarpic dimer (APD), and the like),and/or farnesoid X receptor (FXR) agonists (e.g., GW4064,6alpha-ethyl-chenodeoxycholic acid (6-ECDCA), T0901317, and the like),and/or plasminogen activator-1 (PAI-1) inhibitors (see, e.g.,oxime-based PAI-1 inhibitors, see also U.S. Pat. No. 5,639,726, and thelike), and/or low molecular weight heparin, and/or AGEinhibitors/breakers (e.g., Benfotiamine, aminoguanidine, pyridoxamine,Tenilsetam, Pimagedine, and the like) and/or ADP receptor blockers(e.g., Clopidigrel, AZD6140, and the like), and/or ABCA1 agonists,and/or scavenger receptor B1 agonists, and/or Adiponectic receptoragonist or adiponectin inducers, and/or stearoyl-CoA Desaturase I (SCD1)inhibitors, and/or Cholesterol synthesis inhibitors (non-statins),and/or Diacylglycerol Acyltransferase I (DGAT1) inhibitors, and/orAcetyl CoA Carboxylase 2 inhibitors, and/or LP-PLA2 inhibitors, and/orGLP-1, and/or glucokinase activator, and/or CB-1 agonists, and/oranti-thrombotic/coagulants, and/or Factor Xa inhibitors, and/orGPIIb/IIIa inhibitors, and/or Factor VIIa inhibitors, and/or Tissuefactor inhibitors, and/or anti-inflammatory drugs, and/or Probucol andderivatives (e.g. AGI-1067, etc.), and/or CCR2 antagonists, and/orCX3CR1 antagonists, and/or IL-1 antagonists, and/or nitrates and NOdonors, and/or phosphodiesterase inhibitors, and the like.

In addition, other pharmacologically active materials that can bedelivered in conjunction with one or more of the active agents describedherein include, but are not limited to, agents that reduce the risk ofeye disease events and/or complications thereof. Such agents include,but are not limited to anti-angiogenics or anti-VEGF (anti-VascularEndothelial Growth Factor) agents. For example, the active agentsdescribed herein can be used in conjunction and/or combination withanti-angiogenic therapies for choroidal neovascularization. The neteffect of the combination can result in limiting the expression ofpro-angiogenic factors due to remodeled Bruch's membrane and thus resultin a better metabolic situation for the retinal pigment epithelium andretina, which can limit the duration/circles of anti-angiogenicretreatments.

Angiogenesis is an important cellular event in which vascularendothelial cells proliferate, prune and reorganize to form new vesselsfrom preexisting vascular network. There are compelling evidences thatthe development of a vascular supply is essential for normal andpathological proliferative processes (Folkman and Klagsbrun (1987)Science 235:442-447). Delivery of oxygen and nutrients, as well as theremoval of catabolic products, represent rate-limiting steps in themajority of growth processes occurring in multicellular organisms. Thus,it has been generally assumed that the vascular compartment isnecessary, not only for organ development and differentiation duringembryogenesis, but also for wound healing and reproductive functions inthe adult.

Angiogenesis is also implicated in the pathogenesis of a variety ofdisorders, including but not limited to, tumors, proliferativeretinopathies, age-related macular degeneration, rheumatoid arthritis(RA), and psoriasis. Angiogenesis is essential for the growth of mostprimary tumors and their subsequent metastasis. Tumors can absorbsufficient nutrients and oxygen by simple diffusion up to a size of 1-2mm, at which point their further growth requires the elaboration ofvascular supply. This process is thought to involve recruitment of theneighboring host mature vasculature to begin sprouting new blood vesselcapillaries, which grow towards, and subsequently infiltrate, the tumormass. In addition, tumor angiogenesis involve the recruitment ofcirculating endothelial precursor cells from the bone marrow to promoteneovascularization. Kerbel (2000) Carcinogenesis 21:505-515; Lynden etal. (2001) Nat. Med. 7:1194-1201.

In view of the remarkable physiological and pathological importance ofangiogenesis, much work has been dedicated to the elucidation of thefactors capable of regulating this process. It is suggested that theangiogenesis process is regulated by a balance between pro- andanti-angiogenic molecules, and is derailed in various diseases,especially cancer. Carmeliet and Jain (2000) Nature 407:249-257.

Vascular endothelial cell growth factor (VEGF), which is also termedVEGF-A or vascular permeability factor (VPF), has been reported as apivotal regulator of both normal and abnormal angiogenesis. Ferrara andDavis-Smyth (1997) Endocrine Rev. 18:4-25; Ferrara (1999) J. Mol. Med.77:527-543. Compared to other growth factors that contribute to theprocesses of vascular formation, VEGF is unique in its high specificityfor endothelial cells within the vascular system. VEGF is essential forembryonic vasculogenesis and angiogenesis. (Carmeliet et al. (1996)Nature 380:435-439; Ferrara et al. (1996) Nature 380:439-442).

In addition to being an angiogenic factor in angiogenesis andvasculogenesis, VEGF, as a pleiotropic growth factor, exhibits multiplebiological effects in other physiological processes, such as endothelialcell survival, vessel permeability and vasodilation, monocyte chemotaxisand calcium influx. Ferrara and Davis-Smyth (1997), supra. Moreover,recent studies have reported mitogenic effects of VEGF on a fewnon-endothelial cell types, such as retinal pigment epithelial cells,pancreatic duct cells and Schwann cells. Guerrin et al. (1995) J. CellPhysiol. 164:385-394; Oberg-Welsh et al. (1997) Mol. Cell. Endocrinol.126:125-132; Sondell et al. (1999) J. Neurosci. 19:5731-5740.

Substantial evidence also implicates VEGF's critical role in thedevelopment of conditions or diseases that involve pathologicalangiogenesis. The VEGF mRNA is overexpressed by the majority of humantumors examined (Berkman et al. J Clin Invest 91:153-159 (1993); Brownet al. Human Pathol. 26:86-91 (1995); Brown et al. Cancer Res.53:4727-4735 (1993); Mattern et al. Brit. J. Cancer. 73:931-934 (1996);and Dvorak et al. Am J. Pathol. 146:1029-1039 (1995)). Also, theconcentration of VEGF in eye fluids are highly correlated to thepresence of active proliferation of blood vessels in patients withdiabetic and other ischemia-related retinopathies (Aiello et al. N.Engl. J. Med. 331:1480-1487 (1994)). Furthermore, recent studies havedemonstrated the localization of VEGF in choroidal neovascular membranesin patients affected by AMD (Lopez et al. Invest. Ophtalmo. Vis. Sci.37:855-868 (1996)).

Given its central role in promoting tumor growth, VEGF provides anattractive target for therapeutic intervention. Indeed, a variety oftherapeutic strategies aimed at blocking VEGF or its receptor signalingsystem are currently being developed for the treatment of neoplasticdiseases. Rosen (2000) Oncologist 5:20-27; Ellis et al. (2000)Oncologist 5:11-15; Kerbel (2001) J. Clin. Oncol. 19:45 S-51S. So far,VEGF/VEGF receptor blockade by monoclonal antibodies and inhibition ofreceptor signaling by tyrosine kinase inhibitors are the best studiedapproaches. VEGFR-1 ribozymes, VEGF toxin conjugates, and soluble VEGFreceptors are also being investigated.

Suitable antiangiogenics therefore include, but are not limited topegaptanib (Macugen™ by Pfizer), ranibizumab (Lucentis™ by Genentech)bevacizumab (Avastin™ by Genentech), carboxyamidotriazole, TNP-470,CM101, IFN-α, IL-12, platelet factor 4, suramin, SU5416, thrombospondin,VEGFR antagonists, angiostatic steroids+heparin, cartilage-derivedangiogenesis inhibitory factor, matrix metallopreteinase inhibitors,angiostatin, endostatin, 2-methoxyestradiol, tecogalan, prolactin,α_(v)β₃ inhibitors, and linomide, VEGF-Trap (by RegeneronPharmaceuticals), Aminosterols (Evizion® by Genera Corp.), Cortisen(Retaane® by Alcon), tyrosine kinase inhibitors, anti-angiogenic siRNA,inhibitors of the complement system, and gentherapeutic therapies (e.g.AdPEDF.11 by Genvec).

Other suitable antiangiogenics suitable for the methods described hereinare described, for example, in U.S. Patent Publications 2006/0134111,2007/0031413, 2007/0160608, and the like which are incorporated hereinby reference.

One or more of the active agents can also be administered in conjunctionor combination with compounds that support remodeling of Bruch'smembrane and the adjacent structures (e.g. chelators for iron, calcium,zinc; metalloproteinase inhibitors etc.). As described above, theNational Eye Institute and others have shown that administration ofvitamin supplements with high doses of antioxidants, lutein andzeaxanthin can slow the progression of dry macular degeneration and insome patients, improve visual acuity. As such, one or more of the activeagents can also be administered in conjunction or combination withvitamin supplements with high doses of antioxidants, lutein andzeaxanthin.

One or more of the active agents can also be administered inconjunction, combination, or in a preparation for cell transplants (e.g.stem cells, engineered, autologous, etc.) and biotechnical implantswhere cell survival and outcome of the procedure is improved byremodeled Bruch's membrane and reduced inflammation response.

IX. Kits for the Treatment of One or More Indications

In another embodiment this invention provides kits for amelioration ofone or more symptoms of atherosclerosis or for the prophylactictreatment of a subject (human or animal) at risk for atherosclerosisand/or the treatment or prophylaxis of one or more of the conditionsdescribed herein. For example, also disclosed herein are kits foramelioration of one or more symptoms of atherosclerosis or for theprophylactic treatment of a subject (human or animal) at risk for eyedisease.

The kits preferably comprise a container containing one or more of theactive agents described herein. The active agent(s) can be provided in aunit dosage formulation (e.g. suppository, tablet, caplet, patch, etc.)and/or may be optionally combined with one or more pharmaceuticallyacceptable excipients.

The kit can, optionally, further comprise one or more other agents usedin the treatment of the condition/pathology of interest. Such agentsinclude, but are not limited to, beta blockers, vasodilators, aspirin,statins, ace inhibitors or ace receptor inhibitors (ARBs) and the like,e.g. as described above.

In addition, the kits optionally include labeling and/or instructionalmaterials providing directions (i.e., protocols) for the practice of themethods or use of the “therapeutics” or “prophylactics” of thisinvention. Preferred instructional materials describe the use of one ormore active agent(s) of this invention to mitigate one or more symptomsof atherosclerosis (or other pathologies described herein) and/or toprevent the onset or increase of one or more of such symptoms in anindividual at risk for atherosclerosis (or other pathologies describedherein). The instructional materials may also, optionally, teachpreferred dosages/therapeutic regiment, counter indications and thelike.

While the instructional materials typically comprise written or printedmaterials they are not limited to such. Any medium capable of storingsuch instructions and communicating them to an end user is contemplatedby this invention. Such media include, but are not limited to electronicstorage media (e.g., magnetic discs, tapes, cartridges, chips), opticalmedia (e.g., CD ROM), and the like. Such media may include addresses tointernet sites that provide such instructional materials.

EXAMPLES

The following examples are offered to illustrate, but not to limit theclaimed invention.

Example 1 Use of ApoJ-Related Peptides to Mediate Symptoms ofAtherosclerosis Prevention of LDL-Induced Monocyte Chemotactic Activity

FIG. 1 illustrates a comparison of the effect of D-4F (Anantharamaiah etal. (2002) Circulation, 105: 290-292) with the effect of an apoJ peptidemade from D amino acids (D-J336,Ac-L-L-E-Q-L-N-E-Q-F-N-W-V-S-R-L-A-N-L-T-Q-G-E-NH₂, SEQ ID NO:1177)) onthe prevention of LDL-induced monocyte chemotactic activity in vitro ina co-incubation. Human aortic endothelial cells were incubated withmedium alone (no addition), with control human LDL (200 μg protein/ml)or control human LDL+ control human HDL (350 μg HDL protein/ml). D-J336or D-4F was added to other wells in a concentration range as indicatedplus control human LDL (200 μg protein/ml). Following overnightincubation, the supernatants were assayed for monocyte chemotacticactivity. As shown in FIG. 1, the in vitro concentration of the apoJvariant peptide that prevents LDL-induced monocyte chemotactic activityby human artery wall cells is 10 to 25 times less than the concentrationrequired for the D-4F peptide.

Prevention of LDL-Induced Monocyte Chemotactic Activity by Pre-Treatmentof Artery Wall Cells with D-J336

FIG. 2 illustrates a comparison of the effect of D-4F with the effect ofD-J336 on the prevention of LDL induced monocyte chemotactic activity ina pre-incubation. Human aortic endothelial cells were pre-incubated withD-J336 or D-4F at 4, 2, and 1.mu.g/ml for DJ336 or 100, 50, 25, and 12.5μg/ml for D-4F for 6 hrs. The cultures were then washed and wereincubated with medium alone (no addition), or with control human LDL(200 μg protein/ml), or with control human LDL+ control human HDL (350μg HDL protein/ml) as assay controls. The wells that were pre-treatedwith peptides received the control human LDL at 200 μg protein/ml.Following overnight incubation, the supernatants were assayed formonocyte chemotactic activity.

As illustrated in FIG. 2, the ApoJ variant peptide was 10-25 times morepotent in preventing LDL oxidation by artery wall cells in vitro.

The Effect of Apo J Peptide Mimetics on HDL Protective Capacity in LDLReceptor Null Mice.

D-4F designated as F, or the apoJ peptide made from D amino acids(D-J336, designated as J) was added to the drinking water of LDLreceptor null mice (4 per group) at 0.25 or 0.5 mg per ml of drinkingwater. After 24- or 48-hrs blood was collected from the mice and theirHDL was isolated and tested for its ability to protect againstLDL-induced monocyte chemotactic activity. Assay controls includedculture wells that received no lipoproteins (no addition), or controlhuman LDL alone (designated as LDL, 200.mu.g cholesterol/ml), or controlLDL+ control human HDL (designated as +HDL, 350.mu.g HDL cholesterol).For testing the mouse HDL, the control LDL was added together with mouseHDL (+F HDL or +J HDL) to artery wall cell cultures. The mouse HDL wasadded at 100 μg cholesterol/ml respectively. After treatment with eitherD-4F or D-J336 the mouse HDL at 100 μg/ml was as active as 350 μg/ml ofcontrol human HDL in preventing the control LDL from inducing the arterywall cells to produce monocyte chemotactic activity. The reason for thediscrepancy between the relative doses required for the D-J336 peptiderelative to D-4F in vitro and in vivo may be related to the solubilityof the peptides in water and we believe that when measures are taken toachieve equal solubility the D-J peptides will be much more active invivo as they are in vitro.

Protection Against LDL-Induced Monocyte Chemotactic Activity by HDL fromApo E Null Mice Given Oral Peptides.

FIG. 4 illustrates the effect of oral apoA-1 peptide mimetic and apoJpeptide on HDL protective capacity. ApoE null mice (4 per group) wereprovided with D-4F (designated as F) at 50, 30, 20, 10, 5.mu.g per ml ofdrinking water or apoJ peptide (designated as J) at 50, 30 or 20.mu.gper ml of drinking water. After 24 hrs blood was collected, plasmafractionated by FPLC and fractions containing LDL (designated as mLDLfor murine LDL) and fractions containing HDL (designated as mHDL) wereseparately pooled and HDL protective capacity against LDL oxidation asdetermined by LDL-induced monocyte chemotactic activity was determined.For the assay controls the culture wells received no lipoproteins (noadditions), mLDL alone (at 200 μg cholesterol/ml), or mLDL+ standardnormal human HDL (designated as Cont. h HDL, at 350 μg HDLcholesterol/ml).

For testing the murine HDL, mLDL together with murine HDL (+F mHDL or +JmHDL) were added to artery wall cell cultures. The HDL from the micethat did not receive any peptide in their drinking water is designatedas no peptide mHDL. The murine HDL was used at 100.mu.g cholesterol/ml.After receiving D-4F or D-J336 the murine HDL at 100 μg/ml was as activeas 350.mu.g/ml of normal human HDL. As shown in FIG. 4, when added tothe drinking water the D-J peptide was as potent as D-4F in enhancingHDL protective capacity in apo E null mice.

Ability of LDL Obtained from ApoE Null Mice Given Oral Peptides toInduce Monocyte Chemotactic Activity.

FIG. 5 illustrates the effect of oral apo A-1 peptide mimetic and apoJpeptide on LDL susceptibility to oxidation. ApoE null mice (4 per group)were provided, in their drinking water, with D-4F (designated as F) at50, 30, 20, 10, 5.mu.g per ml of drinking water or the apoJ peptide(D-J336 made from D amino acids and designated as J) at 50, 30 or 20 μgper ml of drinking water. After 24 hrs blood was collected from the miceshown in FIG. 4, plasma fractionated by FPLC and fractions containingLDL (designated as mLDL for murine LDL) were pooled and LDLsusceptibility to oxidation as determined by induction of monocytechemotactic activity was determined. For the assay controls the culturewells received no lipoproteins (no additions), mLDL alone (at 200 μgcholesterol/ml), or mLDL+ standard normal human HDL (designated as Cont.h HDL, 350 μg HDL cholesterol).

Murine LDL, mLDL, from mice that received the D-4F (F mLDL) or thosethat received the apoJ peptide (J mLDL) were added to artery wall cellcultures. LDL from mice that did not receive any peptide in theirdrinking water is designated as No peptide LDL.

As shown in FIG. 5, when added to the drinking water, D-J336 wasslightly more potent than D-4F in rendering the LDL from apo E null miceresistant to oxidation by human artery wall cells as determined by theinduction of monocyte chemotactic activity.

Protection Against Phospholipid Oxidation and Induction of MonocyteChemotactic Activity by HDL Obtained from Apo E Null Mice Given OralPeptides.

FIG. 6 illustrates the effect of oral apoA-1 peptide mimetic and apoJpeptide on HDL protective capacity. ApoE null mice (4 per group) wereprovided with D-4F (designated as F) at 50, 30, 20, 10, 5.mu.g per ml ofdrinking water or apoJ peptide (D-J336 made from D amino acids anddesignated as J) at 50, 30 or 20.mu.g per ml of drinking water. After 24hrs blood was collected, plasma fractionated by FPLC and fractionscontaining HDL (designated as mHDL) were pooled and HDL protectivecapacity against PAPC oxidation as determined by the induction ofmonocyte chemotactic activity was determined. For the assay controls theculture wells received no lipoproteins (no additions), the phospholipidPAPC at 20.mu.g/ml+HPODE, at 1.0 μg/ml, or PAPC+HPODE plus standardnormal human HDL (at 350 μg HDL cholesterol/ml and designated as +Cont.h HDL).

For testing the murine HDL, PAPC+HPODE together with murine HDL (+F mHDLor +J mHDL) were added to artery wall cell cultures. The HDL from micethat did not receive any peptide in their drinking water is designatedas “no peptide mHDL”. The murine HDL was used at 100 μg cholesterol/ml.

The data shown in FIG. 6 indicates that, when added to the drinkingwater, D-J336 was as potent as D-4F in causing HDL to inhibit theoxidation of a phospholipid PAPC by the oxidant HPODE in a human arterywall co-culture as measured by the generation of monocyte chemotacticactivity.

Effect of Oral ApoA-1 Peptide Mimetic and ApoJ Peptide on PlasmaParaoxonase Activity in Mice.

FIG. 7 shows the effect of oral apoA-1 peptide mimetic and apoJ peptideon plasma paraoxonase activity in mice. ApoE null mice (4 per group)were provided with D-4F designated as F at 50, 10, 5 or 0 μg per ml ofdrinking water or apoJ peptide (D-J336 made from D amino acids anddesignated as J) at 50, 10 or 5 μg per ml of drinking water. After 24hrs blood was collected and plasma was assayed for PON1 activity. Thesedata demonstrate that, when added to the drinking water, D-J336 was atleast as potent as D-4F in increasing the paraoxonase activity of apo Enull mice.

Example 2 Oral G* Peptides Increase HDL Protective Capacity in Apo EDeficient Mice

Female, 4 month old apoE deficient mice (n=4 per group) were treatedwith G* peptides having the following amino acid sequences. Peptide113-122=Ac-LVGRQLEEFL-NH₂(SEQ ID NO. 9), Peptide336-357=Ac-LLEQLNEQFNWVSRLANLTQGE-NH₂ (SEQ ID NO. 17), and Peptide377-390=Ac-PSGVTEVVVKLFDS-NH₂ (SEQ ID NO. 19).

Each mouse received 200.mu.g of the peptide by stomach tube. Four hourslater blood was obtained, plasma separated, lipoproteins fractionatedand HDL (at 25 μg per ml) was assayed for protective capacity againstthe oxidation of LDL (at 100 μg per ml) in cultures of human artery wallcells. The data are shown in FIG. 8. The peptide afforded significantHDL-protective capacity in the mice.

In another experiment, female, 4 month old apoE deficient mice (n=4 pergroup) were treated with the 11 amino acid G* peptide 146-156 with thesequence: Ac-QQTHMLDVMQD-NH₂. (SEQ ID NO:11). The mice received thepeptide in their drinking water at the indicated concentrations (seeFIG. 9). Following eighteen hrs, blood was obtained, plasma separated,lipoproteins fractionated and HDL (at 50.mu.g cholesterol per ml) wasassayed for protective capacity against the oxidation of PAPC (at 25 μgper ml)+HPODE (at 1.0 μg per ml) in cultures of human artery wall cells.Assay controls included No additions, PAPC+HPODE and PAPC+HPODE plusControl HDL (designated as +HDL). The data are mean+/−SD of the numberof migrated monocytes in nine high power fields in triplicate cultures.Asterisks indicate significance at the level of p<0.05 vs. the watercontrol (0 μg/ml).

Example 3 Solution Phase Chemistry for Peptide Synthesis

In certain embodiments, a solution-phase synthesis chemistry provides amore economical means of synthesizing peptides of this invention.

Prior to this invention synthesis was typically performed using anall-solid phase synthesis chemistry. The solid phase synthesis ofpeptides of less than 9 amino acids is much more economical than thesolid phase synthesis of peptides of more than 9 amino acids. Synthesisof peptides of more than 9 amino acids results in a significant loss ofmaterial due to the physical dissociation of the elongating amino acidchain from the resin. The solid phase synthesis of peptides containingless than 9 amino acids is much more economical because the there isrelatively little loss of the elongating chain from the resin.

In certain embodiments, the solution phase synthesis functions byconverting the synthesis of the 18 amino acid apoA-I mimetic peptide, 4F(and other related peptides) from an all solid phase synthesis to eitheran all solution phase synthesis or to a combination of solid phasesynthesis of three chains each containing, e.g., 6 amino acids followedby the assembly of the three chains in solution. This provides a muchmore economical overall synthesis. This procedure is readily modifiedwhere the peptides are not 18 amino acids in length. Thus, for example,a 15 mer can be synthesized by solid phase synthesis of three 5 mersfollowed by assembly of the three chains in solution. A 14 mer can besynthesized by the solid phase synthesis of two 5 mers and one 4 merfollowed by assembly of these chains in solution, and so forth.

A) Summary of Synthesis Protocol.

A scheme for the synthesis of the peptide D4F(Ac-D-W-F-K-A-F-Y-D-K-V-A-E-K-F-K-E-A-F-NH₂, (SEQ ID NO:5) isillustrated in Table 20. (The scheme and yields for the synthesis areshown in Table 20.

TABLE 20 Illustrative solution phase synthesis scheme.Methods Used for D4F Synthesis Wt. of Final Wt. Crude Fmoc of PeptideWt. of Pure Amino Coupling Resin (gms) Peptide (mg) Synthesis Resin AcidReagent (gms) Yield (%) Yield (%) Stepwise Rink Amide 6 Equiv HBTU/ 42.0 500 Solid Phase (1 mmole) HOBT 86 25 1.8 gms Stepwise Rink Amide2 Equiv DIC/HOBT 3.9 2.0 450 Solid Phase (1 mmole) 86 22.5 1.8 gmsFragment Rink Amide HBTU/ 3.3 1.0 100 coupling (1 mmole) HOBT 43 10 (6 +6 + 6) 1.8 gms* Synthesis of D4F Fragments Fragments Fragment 1(2HN-KFKEAF (SEQ ID NO: 1178) on rink amide resin(K and E are properly protected) Fragment 2 Cl-TrT-Resin 6 Equiv HBTU/11 2.2 crude 6 residues (5 mmol) HOBT protected stepwise 6.5 gms 32Solid Phase Fmoc-Y(But)-D(But)-K(Boc)-V-A-E(But)-COOH (SEQ ID NO: 1179)Fragment 2 Cl-TrT-Resin 6 Equiv HBTU/ 10 1.8 crude 6 residues (5 mmol)HOBT protected stepwise 6.5 gms 32 Solid PhaseAc-D(But)-W-F-K(Boc)-A-F-COOH (SEQ ID NO: 1180) Synthesis by solutionphase using fragments produced by the solid phase method. Fragment Wangresin. C-terminal hexapeptide (subjected to ammonolysis). Yieldquantitative. 1. NH-K(Boc)-F-K(Boc)-E(But)-A-F-Wang resin (SEQ ID NO:1181) NH-K(Boc)-F-K(Boc)-E(But)-A-F-CO-NH (SEQ ID NO: 1182)? Fragment 2from above was coupled to fragment 1 in DMF using DIC/HOBT.Fmoc-Y(But)-D(But)-K(Bpc)-V-A-E(But)-K(Boc)-F-K(Boc)-E(But)-F-Co-NH2(SEQ ID NO: 1183) 12 residue peptide was characterized as free peptideafter removing protecting groups. Yield was 50% | | Fmoc from theabove-12 rtesidue was removed by piperidine in DMF (20%. After dryingthe peptide was copied to Fragment 3 using DCI/HOBT in DMF.Ac-D(But)-W-F-K(Boc)-A-F-Y(But)-D(but)-K(Boc)-V-A-E(But)-K(Boc)-F-K(Boc)-E(But)-A-FCO-NH (SEQ ID NO: 1184) Protected peptide yield wasquantitative. Protecting groups removed using mixture of TFA (80%),phenol (5%), thioanisole (5%). water)5%), triisopropylsilane (TIS, 5%),stirred for 90 min. Precipitated by ether and purified by C-4 HPLCcolumn. Yield 25%

B) Details of Synthesis Protocol. 1. Fragment Condensation Procedure toSynthesize D-4F

Fragments synthesized for fragment condensation on solid phase are:

Fragment 1: (SEQ ID NO: 1185) Ac-D(OBut)-W-F-K(εBoc)-A-F-COOH;Fragment 2: (SEQ ID NO: 1186)Fmoc-Y(0But)-D(OBut)-K(εBoc)-V-A-E(OBut)-COOH; and Fragment 3(SEQ ID NO: 1187) Fmoc-K(εBoc)F-K(εBoc)-E(OBut)-A-F-Rink amide resin.

Fragment 1 was left on the resin to obtain final peptide amide after TFAtreatment.

To synthesize fragment 1: Fmoc-Phe (1.2 equivalents) was added tochlorotrityl resin (Nova Biochem, 1.3 mMol/g substitution, 5 mMol or 6.5g was used) in presence of six equivalents of DIEA inDMF:dichloromethane (1:1)) and stirred for 4 h. Excess of functionalityon the resin was capped with methanol in presence of dichloromethane andDTEA. After the removal of Fmoc-Fmoc amino acid derivatives (2equivalents) were added using HOBut/HBTU reagents as described above.Final Fmoc-D(OBut)-W-F-K(EBoc)-A-F Chlorotrityl resin was treated withFmoc deblocking agent and acetylated with 6 equivalents of aceticanhydride in presence of diisoprolylethyl amine. The resultingAc-D(OBut)-W-F-K(εBoc)-A-F-resin was treated with a mixture oftrifluoroethanol-acetic acid-dichloromethane (2:2:6, 10 ml/g of resin)for 4 h at room temperature. After removal of the resin by filtration,the solvent was removed by aziotropic distillation with n-hexane undervacuum. The residue (1.8 g) was determined by mass spectral analysis tobe Ac-D(OBut)-W-F-K(εBoc)-A-F-COOH (SEQ ID NO:1188).

Fragment 2, Fmoc-Y(OBut)-D(OBut)-K(εBoc)-V-A-E(OBut)-COOH (SEQ IDNO:1189), was obtained using the procedure described for Fragment 1.Final yield was 2.2 g.

Fragment 3. 0.9 g (0.5 mmol) of Rink amide resin (Nova Biochem) was usedto obtain fragment Rink amide resin was treated with 20% pipetidine indichloromethane for 5 min once and 15 min the second time (Fmocdeblocking reagents). 1.2 equivalents of Fmoc-Phe was condensed usingcondensing agents HOBt/HBTU (2 equivalents in presence of few drops ofdiisopropylethyl amine) (amino acid condensation). Deblocking andcondensation of the rest of the amino acids were continued to obtain theof Fmoc-K(EBoc)F-K(εBoc)-E(OBut)-A-F-rink amide resin (SEQ ID NO:1190).Fmoc was cleaved and the peptide resin K(EBoc)F-K(EBoc)-E(OBut)-A-F-rinkamide resin (SEQ ID NO:1190) was used for fragment condensation asdescribed below.

Fragment 2 in DMF was added to Fragment 3 (1.2 equivalents) usingHOBt-HBTU procedure in presence of DIEA overnight. After washing theresin with DMF and deblocking Fmoc-Fragment 1 (1.2 equivalents) wasadded to the dodecapeptide resin using HOBt-HBTU procedure overnight.

The final peptide resin (3.3 g) was treated with a mixture ofTFA-Phenol-triisopropylsilane-thioanisole-water (80:5:5:5) for 1.5 h (10ml of the reagent/g of the resin). The resin was filtered off and thesolution was diluted with 10 volumes of ether. Precipitated peptide wasisolated by centrifugation and washed twice with ether. 1 g of the crudepeptide was subjected to HPLC purification to obtain 100 mg of thepeptide.

2. Characterization of Peptide.

The peptide was identified by mass spectral and analytical HPLC methods.

As shown in FIG. 14 the product of the solution phase synthesis schemeis very biologically active in producing HDL and pre-beta HDL thatinhibit LDL-induced monocyte chemotaxis in apo E null mice. ApoE nullmice were fed 5 micrograms of the D-4F synthesized as described above(Frgmnt) or the mice were given the same amount of mouse chow withoutD-4F (Chow). Twelve hours after the feeding was started, the mice werebled and their plasma was fractionated on FPLC. LDL (100 microgramsLDL-cholesterol) was added to cocultures of human artery wall cellsalone (LDL) or with a control human HDL (Control HDL) or with HDL (50micrograms HDL-cholesterol) or post-HDL (pHDL; prebeta HDL) from micethat did (Frgmnt) or did not (Chow) receive the D-4F and the monocytechemotactic activity produced was determined.

Example 4 Comparison of D-4F and Reverse (Retro-) D-4F Activity

As shown in FIG. 16, the biological activities of D-4F and reverse RD-4Fare not significantly different. Female apoE null mice were administeredby stomach tube 0, 3, 6, 12, or 25 micrograms of D-4F or Reverse D-4F in100 microliters of water. Blood was obtained 7 hours later and theplasma was fractionated by FPLC. A standard control human LDL was addedto human artery wall cells at a concentration of 100 micrograms ofLDL-cholesterol/mL (LDL). The resulting monocyte chemotactic activitywas normalized to 1.0. The same LDL at the same concentration was addedto the human artery wall cells together with HDL at 50 microgramsHDL-cholesterol/mL from a normal human (hHDL) or from the apoE null micethat received the dose of D-4F or Reverse D-4F shown on the X-axis. Theresulting monocyte chemotactic activity was normalized to that of theLDL added without HDL. The resulting value is the HDL InflammatoryIndex. The results shown are the Mean±S.D. for the data from threeseparate experiments.

Example 5 Effects of L-4F on the Bruch's Membrane of Aged C57Bl/6J-apoENull Mice

Aged C57Bl/6J-apoE null mice are a classic atherosclerosis model withsignificantly elevated plasma cholesterol levels even under standarddiets. Previous studies demonstrated lipid accumulation in Bruch'smembrane mimicking early stages of AMD in these animals (Dithmar et al.(2000) Invest Opthalmol Vis Sci 41:2035-42). An effective clearance ofthis lipid debris via an ApoA-I mimetic peptide (L-4F) could preventearly AMD-like degeneration.

Study Design

L-4F was injected directly into the vitreous cavity of right eyes of 10eight month old C57Bl/6J-apoE null mice. A single dose of 3 μl wasinjected with a L-4F concentration of 400 μg/ml. All left eyes wereuntreated throughout the study and thus be served as controls. Theanimals were then sacrificed 21 days after the injection and the eyeswere immediately enucleated and fixed. Eyes then were processed forelectron microscopy (EM).

Results

EM of 2 mice was reviewed. The treated eyes (L-4F) showed in both casesa significant improvement in Bruch's membrane and RPE morphology as wellas lipid content compared to the untreated eyes (control).

Example 6 Animal Model

Aged C57Bl/6J-apoE null mice are a classic atherosclerosis model withsignificantly elevated plasma cholesterol levels even under standarddiets. Previous studies demonstrated lipid accumulation in Bruch'smembrane mimicking early stages of AMD (Dithmar et al. (2000) InvestOpthalmol Vis Sci 41:2035-42). An effective clearance of this lipiddebris via an ApoA-I/ApoE mimetic peptide will remodel Bruch's membranestructure to a state of wild type animal and restore Bruch's membranefunction with improved hydraulic conductivity and increased metabolicexchange rate. In this study the ApoA-I mimetic peptides L-4F and D-4Fwill be analyzed.

Animals

Female C57Bl/6J-apoE null mice were purchased from Jackson Laboratories(Bar Harbor, Me.). The use of animals was conducted according to theAssociation for Research in Vision and Opthalmology (ARVO) Guidelinesfor the Care and Use of Animals. Animals can be kept in plastic cageswith regular light-dark cycle and will be provided continuous freeaccess to water and food. All animals will receive a regular rodent chowdiet. At 9 months animals will be divided into 8 groups with 7 animalseach.

Single intravitreal injection of 3 μl can be performed in the right eyesof all Study I animals. All left eyes will be kept uninjected and servedas intra-individual negative controls. Group 1 received a concentrationof 200 μg/ml L-4F, group 2 received 400 μg/ml L-4F, group 3 received 800μg/ml L-4F and group 4 served as inter-individual negative control withsham injections (no L-4F).

In addition, single intravitreal injection of 3 μl can be performed inthe right eyes of all Study II animals. All left eyes will be keptuninjected and served as intra-individual negative controls. Group 1received a concentration of 200 μg/ml D-4F, group 2 received 400 μg/mlD-4F, group 3 received 800 μg/ml D-4F and group 4 served asinter-individual negative control with sham injections (no D-4F).

Procedure

Inhalation anesthesia was induced with 5% isofluorane. Additionaltopical 0.5% paracaine anesthesia eye drop were administered as well astropicamide eye drops for pupil dilation. With continuous 3% isofluoraneinhalation anesthesia mice were positioned under a surgery microscope.Eyelids were gently retracted, eyeballs manually protruded and gentlyfixed. A sterile 30½-gauge needle (BectonDickinson, Franklin Lakes,N.J.) were used for sclera penetration and the intraocular position ofthe needle tip was checked under the microscope. A volume of 3 μl ofL-4F or D-4F diluted in saline solution was injected with sterilized 10μl glass micro-syringes (Hamilton, Reno, Nev.) directly into thevitreous cavity. The concentration varied between the 3 groups. Groups 1received 200 μg/ml L-4F (Study I) or D-4F (Study II), group 2 received400 μg/ml L-4F or D-4F and group 3 received saline without L-4F or D-4F.The needle was held in place for 1 min to allow the drug to diffuse intothe vitreous cavity and to prevent retrograde efflux. After theinjection is finished both eyes were treated once with antibiotic 0.3%gentamicin sulphate eye ointment (Gentak®, Akorn Inc., Buffalo Grove,Ill.). After treatment mice were daily observed for adverse events,especially relating to the eye.

After 21 days all animals were sacrificed after deep ketamine/xylazineintraperitoneal anesthesia by thoracotomy/exsanguinations followed by awhole body perfusion with 1.2% paraformaldehyde/0.8% glutaraldehyde in0.1M phosphate buffer. Immediately all right and left eyes wereenucleated and stored in the above mentioned fixative.

Transmission Electron Microscopy

The fixed eyes were bisected under a dissecting scope (SMZ-U, NikonInstruments Inc., Melville N.Y.) for further processing for TEM. Thehalves used for TEM were postfixed in 2% buffered osmium tetroxide,dehydrated in a graded ethanol series, and embedded in epoxy resinaccording to standard protocols. One-gm thick semithin sections werestained with toluidine blue; ultrathin sections will be stained withuranyl acetate and lead citrate and examined with an electron microscope(1200 EXII; JEOL USA, Peabody, Mass.) equipped with a digital camera(Optronics, Goleta, Calif.).

From each specimen, sections including the central retina includingBruch's membrane will be investigated. Images were acquired according toa random sampling procedure using the bars of the supporting grid aspoints of reference, by which 5 consecutive areas adjacent to the rightside of a grid bar will be imaged at a magnification of 5,000× and20,000× and later analyzed.

Image Analysis

The investigator performing electron microscopic image evaluations andmeasurements will be masked regarding the origin of the specimens. WithImageJ (a public domain, Java-based image processing program developedat the National Institutes of Health) a standard grid was placed aboveeach image. For standardization Bruch's membrane will be evaluated withpoint counting stereology only above capillary lumens and not inintercapillary pillar areas.

Statistical Analysis

For statistical analysis of the morphological parameters from TEMimaging, the mean±SD of the individual 30 measurements will becalculated for each eye. The mean values will be compared between groupsusing the non-parametric Mann-Whitney test; differences will beconsidered significant at p<0.05. The SPSS for Windows statisticsprogram (Version 6.0.1, SPSS Inc.) will be used.

Example 7

To study the pharmacokinetics and pharmacodynamics of ApoA-I mimeticpeptides in the eye both eyes of 15 ApoE null mice can be injected with400 μg/ml of biotinylated but functional L-4F or D-4F. Animals can thenbe sacrificed at different time points after injection, 3 mice for eachtime point (1 day, 2 days, 4 days, 7 days, and 12 days after injection).

One eye of each animal can then be cryo-preserved for fluorescentlabeling of the biotinylated compound and light microscopy, the secondeye can then be paraformaldehyde/glutaraldehyde-fixed for electronmicroscopy for evaluation of structural effects as described above.

It is understood that the examples and embodiments described herein arefor illustrative purposes only and that various modifications or changesin light thereof will be suggested to persons skilled in the art and areto be included within the spirit and purview of this application andscope of the appended claims. All publications, patents, and patentapplications cited herein are hereby incorporated by reference in theirentirety for all purposes.

1. A method for treating a subject with eye disease, the methodcomprising administering to the subject in need thereof an effectiveamount of one or more of the active agents described in Tables 2-18,and/or a small organic molecule as described herein in an amountsufficient to ameliorate one or more symptoms of said condition, whereinsaid active agent is administered in combination with a antiangiogenicagents.
 2. The method of claim 1, wherein said active agent is apolypeptide comprising the amino acid sequence of 4F (SEQ ID NO:5). 3.The method of claim 1, wherein said administration is by a routeselected from the group consisting of oral administration, nasaladministration, rectal administration, intraperitoneal injection, andintravascular injection, intraocular injection, intravitreal injection,subconjuctival injection, peri-/retrobulbar injection, subcutaneousinjection, eye drops, eye gel, eye ointment, spray, emulsion,suspension, transcutaneous administration, and intramuscular injection,via any drug carriers as sponges, contact lenses, polymers,microspheres, implants, pellets, and genetically engineered cells. 4.The method of claim 1, wherein the eye disease is selected from thegroup comprising macular degeneration, age related maculopathy (ARM),age related macular degeneration (AMD) including both the dry and wetforms of age related macular degeneration, glaucoma, ocularhypertension, macular edema, retinal pigment epithelium detachments,coats disease, uveitis, sicca syndrome, hereditary diseases associatedwith increased extra-/intracellular lipid storage/accumulation, andjuvenile macular degeneration.
 5. A method of ameliorating a symptom ofeye disease, the method comprising administering to the subject to thesubject in need thereof an effective amount of one or more of the activeagents described in Tables 2-18, and/or a small organic molecule asdescribed herein in an amount sufficient to ameliorate one or moresymptoms of said condition, wherein said active agent is administered incombination with a antiangiogenic agents.
 6. The method of claim 5wherein the symptom is selected from the group comprising accumulationof extracellular lipids in Bruch's membranes, accumulation of lipid richdebris, vision loss, formation of choriocapillaris, thickening of theBruch's membrane, accumulation of neutral lipids in the Bruch'smembrane, formation of a diffusion barrier between the retinal pigmentepithelium and choriocapillaris, deposition of debris (basal lineardeposits and drusen) between the basal membrane of the RPE, and theinner collagenous layer, accumulation of lipofuscin in the RPE cells,RPE atrophy, photoreceptor degeneration, choroidal neovascularization,as well as leakage, bleeding, scarring of the eye.
 7. The method ofclaim 5, wherein the eye disease is selected from the group comprisingmacular degeneration, age related maculopathy (ARM), age related maculardegeneration (AMD) including both the dry and wet forms of age relatedmacular degeneration, glaucoma, ocular hypertension, macular edema,retinal pigment epithelium detachments, coats disease, uveitis, siccasyndrome, hereditary diseases associated with increasedextra-/intracellular lipid storage/accumulation, and juvenile maculardegeneration,
 8. A method of ameliorating a symptom of eye disease, themethod comprising administering to the subject an effective amount of apeptide wherein said peptide: ranges in length from about 18 to 37 aminoacids and comprises at least 3 alanines (A), 2 aspartates (D), 2glutamates (E), 4 phenylalanines (F), 4 lysines (K), 1 valine (V), 1tryptophan (W), 1 tyrosine (Y); wherein said peptide forms a class Aamphipathic helix; comprises at least one “D” amino acid residue; andprotects a phospholipid against oxidation by an oxidizing agent, whereinsaid active agent is administered in combination with a antiangiogenicagents.
 9. The method of claim 8, wherein said peptide further comprisesa protecting group coupled to the amino or carboxyl terminus.
 10. Themethod of claim 8, wherein said peptide further comprises a firstprotecting group coupled to the amino terminus and a second protectinggroup coupled to the carboxyl terminus.
 11. The method of claim 9,wherein said protecting groups are independently selected from the groupconsisting of acetyl, amide, and 3 to 20 carbon alkyl groups, Fmoc,Tboc, 9-fluoreneacetyl group, 1-fluorenecarboxylic group,9-florenecarboxylic group, 9-fluorenone-1-carboxylic group,benzyloxycarbonyl, Xanthyl (Xan), Trityl (Trt), 4-methyltrityl (Mtt),4-methoxytrityl (Mmt), 4-methoxy-2,3,6-trimethyl-benzenesulphonyl (Mtr),Mesitylene-2-sulphonyl (Mts), 4,4-dimethoxybenzhydryl (Mbh), Tosyl(Tos), 2,2,5,7,8-pentamethyl chroman-6-sulphonyl (Pmc), 4-methylbenzyl(MeBzl), 4-methoxybenzyl (MeOBzl), Benzyloxy (BzlO), Benzyl (Bzl),Benzoyl (Bz), 3-nitro-2-pyridinesulphenyl (Npys),1-(4,4-dimentyl-2,6-diaxocyclohexylidene)ethyl (Dde), 2,6-dichlorobenzyl(2,6-DiCl-Bzl), 2-chlorobenzyloxycarbonyl(2—Cl-Z),2-bromobenzyloxycarbonyl (2—Br-Z), Benzyloxymethyl (Born),t-butoxycarbonyl (Boc), cyclohexyloxy (cHxO), t-butoxymethyl (Bum),t-butoxy (tBuO), t-Butyl (tBu), Acetyl (Ac), and Trifluoroacetyl (TFA).12. The method of claim 8, wherein all enantiomeric amino acids are “D”amino acids.
 13. The method of claim 8, wherein said peptide is mixedwith a pharmacologically acceptable excipient.
 14. The method of claim8, wherein said peptide is mixed with a pharmacologically acceptableexcipient suitable for oral administration to a mammal.
 15. The methodof claim 8, wherein said peptide comprises an amino acid sequenceselected from the group consisting ofD-W-F-K-A-F-Y-D-K-V-A-E-K-F-K-E-A-F (SEQ ID NO: 1191),-D-W-L-K-A-F-Y-D-K-V-A-E-K-L-K-E-A-F-P-D-W-L-K-A-F-Y-D-K-V-A-E-K-L-K-E-A-F(SEQ ID NO: 1192),-D-W-L-K-A-F-Y-D-K-V-A-E-K-L-K-E-F-F-P-D-W-L-K-A-F-Y-D-K-V-A-E-K-L-K-E-F-F(SEQ ID NO: 1193),-D-W-F-K-A-F-Y-D-K-V-A-E-K-L-K-E-A-F-P-D-W-F-K-A-F-Y-D-K-V-A-E-K-L-K-E-A-F(SEQ ID NO: 1194),D-K-L-K-A-F-Y-D-K-V-F-E-W-A-K-E-A-F-P-D-K-L-K-A-F-Y-D-K-V-F-E-W-L-K-E-A-F(SEQ ID NO: 1195),D-K-W-K-A-V-Y-D-K-F-A-E-A-F-K-E-F-L-P-D-K-W-K-A-V-Y-D-K-F-A-E-A-F-K-E-F-L(SEQ ID NO: 1196),D-W-F-K-A-F-Y-D-K-V-A-E-K-F-K-E-A-F-P-D-W-F-K-A-F-Y-D-K-V-A-E-K-F-K-E-A-F-(SEQID NO: 1197), or the reverse of any of these sequences.
 16. The methodof claim 15, wherein said peptide comprises a protecting group coupledto the amino terminal and said amino terminal protecting group is aprotecting group selected from the group consisting of acetyl,propeonyl, and a 3 to 20 carbon alkyl.
 17. The method of claim 15,wherein said peptide comprises a protecting group coupled to thecarboxyl terminal and said carboxyl terminal protecting group is anamide.
 18. The method of claim 15, wherein said peptide comprises: afirst protecting group coupled to the amino terminus wherein saidprotecting group is a protecting group selected from the groupconsisting of acetyl, propeonyl, and a 3 to 20 carbon alkyl; and asecond protecting group coupled to the carboxyl terminal and saidcarboxyl terminal protecting group is an amide.
 19. The method of claim8, wherein said oxidizing agent is selected from the group consisting ofhydrogen peroxide, 13(S)-HPODE, 15(S)-HPETE, HPODE, HPETE, HODE, andHETE.
 20. The method of claim 8, wherein said phospholipid is selectedfrom the group consisting of1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphorylcholine (Ox-PAPC),1-palmitoyl-2-oxovaleroyl-sn-glycero-3-phosphorylcholine (POVPC),1-palmitoyl-2-glutaroyl-sn-glycero-3-phosphorylcholine (PGPC),1-palmitoyl-2-epoxyisoprostane-sn-glycero-3-phosphorylcholine (PEIPC),1-stearoyl-2-arachidonoyl-sn-glycero-3-phosphorylcholine (SAPC),1-stearoyl-2-oxovaleroyl-sn-glycero-3-phosphorylcholine (SOVPC),1-stearoyl-2-glutaroyl-sn-glycero-3-phosphorylcholine (SGPC),1-stearoyl-2-epoxyisoprostane-sn-glycero-3-phosphorylcholine (SEIPC),1-stearoyl-2-arachidonyl-sn-glycero-3-phosphorylethanolamine(Ox-SAPE),1-stearoyl-2-oxovaleroyl-sn-glycero-3-phosphorylethanolamine(SOVPE),1-stearoyl-2-glutaroyl-sn-glycero-3-phosphorylethanolamine(SGPE), and1-stearoyl-2-epoxyisoprostane-sn-glycero-3-phosphorylethanolamine(SEIPE).
 21. The method of claim 1, wherein the anti-angiogenic therapyis selected from the list consisting of pegaptanib (Macugen™ by Pfizer),ranibizumab (Lucentis™ by Genentech) bevacizumab (Avastin™ byGenentech), carboxyamidotriazole, TNP-470, CM101, IFN-α, IL-12, plateletfactor 4, suramin, SU5416, thrombospondin, VEGFR antagonists,angiostatic steroids+heparin, cartilage-derived angiogenesis inhibitoryfactor, matrix metallopreteinase inhibitors, angiostatin, endostatin,2-methoxyestradiol, tecogalan, prolactin, α_(v)β₃ inhibitors, andlinomide, VEGF-Trap (by Regeneron Pharmaceuticals), Aminosterols(Evizion® by Genera Corp.), Cortisen (Retaane® by Alcon), tyrosinekinase inhibitors, anti-angiogenic siRNA, inhibitors of the complementsystem, gentherapeutic therapies (e.g. AdPEDF.11 by Genvec).
 22. Themethod of claim 5, wherein the anti-angiogenic therapy is selected fromthe list consisting of pegaptanib (Macugen™ by Pfizer), ranibizumab(Lucentis™ by Genentech) bevacizumab (Avastin™ by Genentech),carboxyamidotriazole, TNP-470, CM101, IFN-α, IL-12, platelet factor 4,suramin, SU5416, thrombospondin, VEGFR antagonists, angiostaticsteroids+heparin, cartilage-derived angiogenesis inhibitory factor,matrix metallopreteinase inhibitors, angiostatin, endostatin,2-methoxyestradiol, tecogalan, prolactin, α_(v)β₃ inhibitors, andlinomide., VEGF-Trap (by Regeneron Pharmaceuticals), Aminosterols(Evizion® by Genera Corp.), Cortisen (Retaane® by Alcon), tyrosinekinase inhibitors, anti-angiogenic siRNA, inhibitors of the complementsystem, gentherapeutic therapies (e.g. AdPEDF.11 by Genvec).